Macrocyclic compounds

ABSTRACT

The present invention relates to macrocyclic compounds which are capable of selective binding to a target saccharide (e.g. glucose), making them particularly well suited for use in saccharide sensing applications. The present invention also relates to processes for the preparation of said compounds, to compositions and devices comprising them, and to their use in the detection of a target saccharide.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. Pat. Application 16/993,772,filed Aug. 14, 2020, which is a continuation of U.S. Pat. Application16/491,379, filed Sep. 5, 2019, (now U.S. Pat. 10,800,747, issued Oct.13, 2020), which is a 35 U.S.C. §371 National Stage application ofInternational Application PCT/GB2018/050679 (WO 2018/167503), filed Mar.15, 2018, which claims priority to Great Britain Patent Applications1704125.2, filed Mar. 15, 2017, and 1715210.9, filed Sep. 20, 2017; thecontents of which are incorporated herein by reference.

INTRODUCTION

The present invention relates to macrocyclic compounds which are capableof selectively binding to a target saccharide (e.g. glucose), makingthem particularly well suited for use in saccharide sensingapplications. The present invention also relates to processes for thepreparation of said compounds, to compositions and devices comprisingthem, and to their use in the detection of a target saccharide.

BACKGROUND OF THE INVENTION

The detection and subsequent monitoring of saccharides, in particularglucose, has many practical applications in both medical and non-medicalapplications. For instance, the reliable detection of glucose in anindividual’s bloodstream underpins the vast majority of treatmentscurrently available for the treatment of diabetes, a condition which theWorld Health Organisation (WHO) state affected around 422 million in2014, and which it projects to be the 7th leading cause of death by2030.

Diabetes, however, is not the only practical application for saccharidedetection, the accurate determination of sugar levels in fermentationmedia, for example, in brewing processes and/or cell culturing, is alsohighly desired. Here, the ability to closely monitor the precise levelsof sugars present during fermentation can be hugely advantageous infine-tuning both the yield and properties of the end product.

Detection and subsequent monitoring of saccharides, however, is heavilyreliant on the provision of saccharide receptor molecules which arecapable of binding to, and thus detecting, saccharides in the aqueousmedia in which they are typically found, so called ‘synthetic lectins’.However, historically, the binding of saccharides in aqueous media hasproven to be a very challenging task for synthetic chemists, and evennatural carbohydrate-binding proteins, known as lectins, struggle todisplay binding affinities in the order of magnitude commonly found innature for such protein-substrate binding interactions.

Saccharides are hydrophilic species, often bearing hydromimetic hydroxylgroups, which makes them well hydrated and of significant resemblance towater molecules. In this regard, successful binding requires a receptormolecule to be able to distinguish between the hydroxyl groups of thesaccharide and an array of water molecules, which are usually present ina much higher abundance than the target saccharide. Furthermore, forbinding to occur, water must be displaced from both the receptormolecule and the saccharide, for which the energetic consequences areoften difficult to predict, thereby making the modelling and design ofsuch receptor molecules difficult.

Moreover, the ability to selectively target one saccharide molecule overother saccharide molecules also presents a significant challenge. Forinstance, for specific saccharide binding to occur, the receptormolecule must be able to distinguish between numerous saccharidemolecules often bearing only very subtle structural differences (i.e.the configuration of a single asymmetric centre).

Despite the above challeneges, a few successful synthetic saccharidereceptor molecules, which are capable of selective sacchariderecognition in aqueous media, have been reported (see, for example,WO2013160701). However, to further advance the use of such syntheticsaccharide receptors in saccharide detection applications, such as thoseoutlined above, there remains a need for new and improved receptormolecules which are capable of displaying higher affinities and/orselectivities towards specific target saccharides (e.g. glucose).

The present invention was devised with the foregoing in mind.

SUMMARY OF THE INVENTION

According to one aspect of the present invention, there is provided acompound, or a salt, hydrate or solvate thereof, as defined herein.

According to a second aspect of the present invention, there is provideda complex comprising a compound, or a salt, hydrate or solvate thereof,as defined herein, in association with a target saccharide.

According to a third aspect of the present invention, there is provideda complex comprising a compound, or a salt, hydrate or solvate thereof,as defined herein, in association with a displaceable reporter molecule.

According to a fourth aspect of the present invention, there is provideda composition comprising a compound, or a salt, hydrate or solvatethereof, as defined herein, and a displaceable reporter molecule.

According to a fifth aspect of the present invention there is provided asaccharide detection device comprising a complex, as defined herein, acomposition, as defined herein, or a compound as defined herein.

According to another aspect of the present invention, there is provideda use of a complex, as defined herein, a composition, as defined herein,a saccharide detection device, as defined herein, or a compound, asdefined herein, for detecting a target saccharide in an aqueousenvironment.

According to a further aspect of the present invention, there isprovided a kit comprising a compound, as defined herein, and adisplaceable reporter molecule.

According to a further aspect of the present invention, there isprovided a process for preparing a compound, or a salt, hydrate orsolvate thereof, as defined herein.

According to yet a further aspect of the present invention, there areprovided novel intermediates as defined herein which are suitable foruse in any one of the synthetic methods set out herein.

Features, including optional, suitable, and preferred features inrelation to one aspect of the invention may also be features, includingoptional, suitable and preferred features in relation to any otheraspect of the invention.

BRIEF DESCRIPTION OF THE DRAWINGS

The patent or application file contains at least one drawing executed incolor. Copies of this patent or patent application publication withcolor drawing(s) will be provided by the Office upon request and paymentof the necessary fee.

FIGS. 1A-1D shows schematic illustations of the key interactions madebetween the target saccharide and the compounds of the present invention(FIGS. 1A and 1B), as well as molecular models of a ground stateconformation of one particular compound of the present invention withglucose (FIGS. 1C and 1D). In FIG. 1C, ten intermolecular NH⋯O hydrogenbonds (with a distance of between 1.9 and 2.2 Å) can be seen, and FIG. 1d further depicts the close CH-π contacts made between the saccharideand compound of the present invention.

FIGS. 2A-2B shows: 2A) the partial ¹H NMR spectra; and 2B) the bindinganalysis curve for receptor 1 (0.25 mM) titrated with a combinedsolution of D-glucose (9.6 mM) and receptor 1 (0.25 mM), in D₂O bufferedwith 10 mM phosphate buffer solution (pH 7.4) at 298 K. Spectra implybinding with slow exchange on NMR timescale. Integrations of peak at8.04 ppm (denoted with •) versus region 8.22-7.21 ppm were plottedagainst D-glucose concentration (mM). The calculated values for theintegrals are overlaid with the observed values, giving K_(a) = 18,026 ±208 M⁻¹ (1.04%).

FIG. 3 shows the ¹H NMR spectra for receptor 1 (0.25 mM) titrated with acombined solution of D-glucose (9.6 mM) and receptor 1 (0.25 mM), in D₂Obuffered with 10 mM phosphate buffer solution (pH 7.4) at 298 K.

FIGS. 4A-4B shows the ¹H NMR spectra at certain time intervals of: a)α-D-glucose (5 mM, and b) α-D-glucose (5 mM) with receptor 1 (0.2 mM).Relative integrals of α-H1 (5.22 ppm) and β-H2 (3.23 ppm) protons overtime were calculated to determine if receptor 1 affected the rate ofanomerisation between α and β-D-glucose. Rate of anomerisation was foundto be independent of receptor 1 (see Table 2).

FIG. 5 shows a plot of relative integrals of αH1:βH2 versus time (min).Similar gradients suggest receptor does not affect the rate ofanomerisation of D-glucose.

FIGS. 6A-6D shows the ITC binding results for receptor 1 (0.13 mM)titrated with glucose (7.5 mM) in H₂O, in which: 6A) shows the blank ITCrun (addition of sugar into water); 6B) shows the actual run (sugar intoreceptor 1); 6C) shows the plotted change in enthalpy vs molar ratio andthe fit calculated with the supplied ITC software (K_(a) = 21,000 ± 2640M⁻¹); and 6D) shows the fit calculated using an Excel spreadsheet tocorroborate the result.

FIGS. 7A-7D shows the ITC binding results for receptor 1 (0.06 mM)titrated with D-glucose (7 mM) in 10 mM PBS buffer (pH 7.4), in which:7A) shows the blank ITC run (addition of sugar into water); 7B) showsthe actual run (sugar into receptor 1); 7C) shows the plotted change inenthalpy vs molar ratio and the fit calculated with the supplied ITCsoftware (K_(a) = 19,100 ± 1310 M⁻¹); and 7D) shows the fit calculatedusing an Excel spreadsheet to corroborate the result.

FIGS. 8A-8D shows the ITC binding results for receptor 1 (0.06 mM)titrated with D-glucose (7 mM) in 10 mM PBS buffer (pH 6), in which: 8A)shows the blank ITC run (addition of sugar into water); 8B) shows theactual run (sugar into receptor 1); 8C) shows the plotted change inenthalpy vs molar ratio and the fit calculated with the supplied ITCsoftware (K_(a) = 19,800 ± 1290 M⁻¹); and 8D) shows the fit calculatedusing an Excel spreadsheet to corroborate the result.

FIGS. 9A-9D shows the ITC binding results for receptor 1 (0.06 mM)titrated with D-glucose (7 mM) in 10 mM PBS buffer (pH 8), in which: 9A)shows the blank ITC run (addition of sugar into water); 9B) shows theactual run (sugar into receptor 1); 9C) shows the plotted change inenthalpy vs molar ratio and the fit calculated with the supplied ITCsoftware (K_(a) = 23,400 ± 1850 M⁻¹); and 9D) shows the fit calculatedusing an Excel spreadsheet to corroborate the result.

FIGS. 10A-10D shows the ITC binding results for receptor 1 (0.06 mM)titrated with D-glucose (7 mM) in DMEM Cell Culture Medium (no glucose,10k MWCO, 90% v/v) and 10 mM phosphate buffer solution (pH 7.4), inwhich: 10A) shows the blank ITC run (addition of sugar into medium);10B) shows the actual run (sugar into receptor 1); 10C) shows theplotted change in enthalpy vs molar ratio; and 10D) shows the fitcalculated using an Excel spreadsheet (Ka = 5637 ± 118 M⁻¹).

FIGS. 11A-11D shows the ITC binding results for receptor 1 (0.06 mM)titrated with D-glucose (7 mM) in 10 mM phosphate buffer solution (pH7.4) with added salts: ferric nitrate (0.2 µM), calcium chloride (1.8mM), magnesium sulfate (0.81 mM), potassium chloride (5.3 mM), sodiumbicarbonate (44 mM), sodium chloride (110 mM) and sodium phosphatemonobasic (0.9 mM), in which: 11A) shows the blank ITC run (addition ofsubstrate into medium); 11B) shows the actual run (substrate intoreceptor 1); 11C) shows the plotted change in enthalpy vs molar ratio;and 11D) shows the fit calculated using an Excel spreadsheet (Ka = 5164± 303 M⁻¹).

FIG. 12 shows ¹H NMR spectra showing receptor 1 (0.1 mM) dissolved inD₂O with 10 mM phosphate buffer (pH 7.4) at 298 K. Addition of MgSO₄(0.8 mM) and CaCl₂ (1.8 mM), which are the concentrations present inDMEM cell culture media, to free receptor showed a small change inchemical shift (δ in ppm) for proton s2. Addition of 2 equivalents (0.2mM) of D-glucose did not saturate the receptor. Addition of this sameconcentration of glucose to free receptor in D₂O with no added salts(top spectrum) did saturate the receptor, suggesting that Ca²⁺ and Mg²⁺inhibit binding.

FIGS. 13A-13D shows the ITC binding results for receptor 1 (0.06 mM)titrated with D-glucose (7 mM) in Leibovitz’s L-15 Cell Culture Medium(no glucose, 10k MWCO, 90% v/v) and 10 mM phosphate buffer solution (pH7.4), in which: 13A) shows the blank ITC run (addition of sugar intomedium); 13B) shows the actual run (sugar into receptor 1); 13C) showsthe plotted change in enthalpy vs molar ratio; and 13D) shows the fitcalculated using an Excel spreadsheet (Ka = 5214 ± 452 M⁻¹).

FIGS. 14A-14D shows the ITC binding results for receptor 1 (0.06 mM)titrated with D-glucose (5 mM) in Human Blood Serum (no glucose, 10kMWCO, 90% v/v) and 10 mM phosphate buffer solution (pH 8.5), in which:14A) shows the blank ITC run (addition of sugar into medium); 14B) showsthe actual run (sugar into receptor 1); 14C) shows the plotted change inenthalpy vs molar ratio; and 14D) shows the fit calculated using anExcel spreadsheet (Ka = 2477 ± 142 M⁻ ¹).

FIGS. 15A-15B shows: 15A) the ¹H NMR spectra; and 15B) the bindinganalysis curve for receptor 1 (0.07 mM) titrated with a combinedsolution of D-methyl-β-glucoside (10 mM) and receptor 1 (0.07 mM), inD₂O buffered with 10 mM phosphate buffer solution (pH 7.4) at 298 K.Spectra imply binding with slow exchange on NMR timescale. Integrationsof peak at 8.31 ppm (denoted with •) versus region 8.36-7.36 ppm wereplotted against guest concentration (mM). The calculated values for theintegrals are overlaid with the observed values, giving K_(a) = 7522 ±414 M⁻¹ (5.51%).

FIGS. 16A-16D shows the ITC binding results for receptor 1 (0.13 mM)titrated with methyl-β-D-glucoside (7 mM) in H₂O, in which: 16A) showsthe blank ITC run (addition of sugar into water); 16B) shows the actualrun (sugar into receptor 1); 16C) shows the plotted change in enthalpyvs molar ratio and the fit calculated with the supplied ITC software(K_(a) = 9120 ± 542 M⁻ ¹); and 16D) shows the fit calculated using anExcel spreadsheet to corroborate the result.

FIGS. 17A-17D shows the ITC binding results for receptor 1 (0.06 mM)titrated with methyl-β-D-glucoside (7 mM) in 10 mM phosphate buffersolution (pH 7.4), in which: 17A) shows the blank ITC run (addition ofsugar into water); 17B) shows the actual run (sugar into receptor 1);17C) shows the plotted change in enthalpy vs molar ratio; and 17D) showsthe fit calculated using an Excel spreadsheet (K_(a) = 7886 ± 1296 M⁻¹).

FIGS. 18A-18D shows the ITC binding results for receptor 1 (0.1 mM)titrated with D-glucuronic acid (5 mM) in 10 mM phosphate buffersolution (pH 7.4) in which: 18A) shows the blank ITC run (addition ofsubstrate into medium); 18B) shows the actual run (substrate intoreceptor 1); 18C) shows the plotted change in enthalpy vs molar ratio;and 18D) shows the fit calculated using an Excel spreadsheet (Ka = 5348± 189 M⁻¹).

FIG. 19 shows the ¹H NMR spectra for receptor 1 (0.1 mM) titrated with acombined solution of D-gluconate (10 mM) and receptor 1 (0.1 mM), in D₂Obuffered with 10 mM phosphate buffer solution (pH 7.4) at 298 K. Spectraimply no binding was observed, despite some broadening of peaks at highconcentrations of guest.

FIGS. 20A-20C shows the ITC binding results for receptor 1 (0.06 mM)titrated with Glucono-δ-lactone/gluconic acid (200 mM) in 10 mMphosphate buffer solution (pH 7.4), in which: 20A) shows the blank ITCrun (addition of substrate into water); 20B) shows the actual run(substrate into receptor 1); 20C) shows the plotted change in enthalpyvs molar ratio.

FIGS. 21A-21B shows: 21A) the partial ¹H NMR spectra; and 21B) thebinding analysis curve for receptor 1 (0.05 mM) titrated with a combinedsolution of D-galactose (250 mM) and receptor 1 (0.05 mM), in D₂Obuffered with 10 mM phosphate buffer solution (pH 7.4) at 298 K. Spectraimply binding with fast/intermediate exchange on NMR timescale. Changesin chemical shift (Δδ ppm) of peak at 7.63 ppm (denoted with •) wereplotted against increasing guest concentration (mM). The calculatedvalues for the Δδ are overlaid with the observed values giving K_(a) =132 ± 13 M⁻¹ (10.2%).

FIGS. 22A-22C shows for receptor 1 (0.06 mM) titrated with D-galactose(518 mM) in H₂O, in which: 22A) shows the blank ITC run (addition ofsugar into water); 22B) shows the actual run (sugar into receptor 1);and 22C) shows the plotted change in enthalpy vs molar ratio.

FIGS. yre 23A-23D shows the shows the ITC binding results for receptor 1(0.1 mM) titrated with D-galactose (75 mM) in 10 mM phosphate buffersolution (pH 7.4) in which: 23A) shows the blank ITC run (addition ofsubstrate into medium); 23B) shows the actual run (substrate intoreceptor 1); 23C) shows the plotted change in enthalpy vs molar ratio;and 23D) shows the fit calculated using an Excel spreadsheet (Ka = 182 ±4.2 M⁻¹).

FIG. 24 shows the partial ¹H NMR spectra for receptor 1 (0.1 mM)titrated with a combined solution of 2-deoxy-D-glucose (50 mM) andreceptor 1 (0.1 mM), in D₂O buffered with 10 mM phosphate buffersolution (pH 7.4) at 298 K. Spectra imply binding with intermediate rateof exchange (rate between fast and slow exchange rates between H and HGspecies) on NMR timescale. Due to severe broadening of peaks forreceptor 1 upon addition of guest, no K_(a) was determinable.

FIGS. 25A-25D shows the ITC binding results for receptor 1 (0.06 mM)titrated with 2-deoxy-D-glucose (7 mM) in H₂O, in which: 25A) shows theblank ITC run (addition of sugar into water); 25B) shows the actual run(sugar into receptor 1); 25C) shows the plotted change in enthalpy vsmolar ratio and the fit calculated with the supplied ITC software (K_(a)= 657 ± 90 M⁻¹); and 25D) shows the fit calculated using an Excelspreadsheet to corroborate the result.

FIGS. 26A-26D shows the ITC binding results for receptor 1 (0.06 mM)titrated with 2-deoxy-D-glucose (7 mM) in 10 mM phosphate buffersolution (pH 7.4), in which: 26A) shows the blank ITC run (addition ofsugar into water); 26B) shows the actual run (sugar into receptor 1);26C) shows the plotted change in enthalpy vs molar ratio; and 26D) showsthe fit calculated using an Excel spreadsheet (Ka = 725 ± 41 M⁻¹).

FIGS. 27A-27B shows: 27A) the partial ¹H NMR spectra; and 27B) thebinding analysis curve for receptor 1 (0.11 mM) titrated with a combinedsolution of D-mannose (250 mM) and receptor 1 (0.11 mM), in D₂O bufferedwith 10 mM phosphate buffer solution (pH 7.4) at 298 K. Spectra implybinding with fast/intermediate exchange on NMR timescale. Changes inchemical shift (Δδ ppm) of peak at 7.63 ppm (denoted with •) wereplotted against increasing guest concentration (mM). The calculatedvalues for the Δδ are overlaid with the observed values giving K_(a) =140 ± 2 M⁻¹ (1.31%).

FIGS. 28A-28C shows for receptor 1 (0.06 mM) titrated with D-Mannose(504 mM) in H₂O, in which: 28A) shows the blank ITC run (addition ofsugar into water); 28B) shows the actual run (sugar into receptor 1);and 29C) shows the plotted change in enthalpy vs molar ratio.

FIGS. 29A-29D shows the ITC binding results for receptor 1 (0.1 mM)titrated with D-mannose (75 mM) in 10 mM phosphate buffer solution (pH7.4) in which: 29A) shows the blank ITC run (addition of substrate intomedium); 29B) shows the actual run (substrate into receptor 1XX); 29C)shows the plotted change in enthalpy vs molar ratio; and 29D) shows thefit calculated using an Excel spreadsheet (K_(a) = 143 ± 1.5 M⁻¹).

FIGS. 30A-30D shows the ITC binding results for receptor 1 (0.1 mM)titrated with D-xylose (5 mM) in 10 mM phosphate buffer solution (pH7.4) in which: 30A) shows the blank ITC run (addition of substrate intomedium); 30B) shows the actual run (substrate into receptor 1); 30C)shows the plotted change in enthalpy vs molar ratio; and 30D) shows thefit calculated using an Excel spreadsheet (K_(a) = 5804 ± 174 M⁻¹).

FIG. 31 shows the partial ¹H NMR spectra for receptor 1 (0.11 mM)titrated with a combined solution of D-cellobiose (250 mM) and receptor1 (0.11 mM), in D₂O buffered with 10 mM phosphate buffer solution (pH7.4) at 298 K. Spectra imply binding with slow exchange on NMRtimescale. Integrations of peak at 8.02 ppm (denoted with •) versusregion 8.36-7.36 ppm were used to calculate the K_(a) (M⁻¹) at eachpoint of addition (see Table 3), an average of these calculated valuesgives K_(a) = 31 ± 2.66 (9%).

FIGS. 32A-32D shows the ITC binding results for receptor 1 (0.06 mM)titrated with cellobiose (250 mM) in H₂O, in which: 32A) shows the blankITC run (addition of sugar into water); 32B) shows the actual run (sugarinto receptor 1); 32C) shows the plotted change in enthalpy vs molarratio and the fit calculated with the supplied ITC software (K_(a) =36.6 ± 2.5 M⁻¹); and 32D) shows the fit calculated using an Excelspreadsheet to corroborate the result

FIGS. 33A-33D shows the ITC binding results for receptor 1 (0.6 mM)titrated with D-cellobiose (250 mM) in 10 mM phosphate buffer solution(pH 7.4) in which: 33A) shows the blank ITC run (addition of substrateinto medium); 33B) shows the actual run (substrate into receptor 1);33C) shows the plotted change in enthalpy vs molar ratio; and 33D) showsthe fit calculated using an Excel spreadsheet (K_(a) = 30.9 ± 4.9 M⁻¹).

FIGS. 34A-34B shows: 34A) the partial ¹H NMR spectra; and 34B) thebinding analysis curve for receptor 1 (0.11 mM) titrated with a combinedsolution of D-fructose (250 mM) and receptor 1 (0.11 mM), in D₂Obuffered with 10 mM phosphate buffer solution (pH 7.4) at 298 K. Spectraimply binding with fast/intermediate exchange on NMR timescale. Changesin chemical shift (Δδ ppm) of peak at 7.63 ppm (denoted with •) wereplotted against increasing guest concentration (mM). The calculatedvalues for the Δδ are overlaid with the observed values giving K_(a) =51 ± 3 M⁻¹ (5.46%).

FIGS. 35A-35D shows the ITC binding results for receptor 1 (0.1 mM)titrated with D-fructose (75 mM) in 10 mM phosphate buffer solution (pH7.4) in which: 35A) shows the blank ITC run (addition of substrate intomedium); 35B) shows the actual run (substrate into receptor 1); 35C)shows the plotted change in enthalpy vs molar ratio; and 35D) shows thefit calculated using an Excel spreadsheet (K_(a) = 60.3 ± 1.6 M⁻¹).

FIGS. 36A-36B shows: 36A) the partial ¹H NMR spectra; and 36B) thebinding analysis curve for receptor 1 (0.11 mM) titrated with a combinedsolution of D-ribose (250 mM) and receptor 1 (0.11 mM), in D₂O bufferedwith 10 mM phosphate buffer solution (pH 7.4) at 298 K. Spectra implybinding with fast exchange on NMR timescale. Changes in chemical shift(Δδ ppm) of peak at 7.83 ppm (denoted with •) were plotted againstincreasing guest concentration (mM). The calculated values for the Δδare overlaid with the observed values giving K_(a) = 264 ± 10 M⁻¹(3.96%).

FIGS. 37A-37D shows the ITC binding results for receptor 1 (0.1 mM)titrated with D-ribose (75 mM) in 10 mM phosphate buffer solution (pH7.4) in which: 37A) shows the blank ITC run (addition of substrate intomedium); 37B) shows the actual run (substrate into receptor 1); 37C)shows the plotted change in enthalpy vs molar ratio; and 37D) shows thefit calculated using an Excel spreadsheet (K_(a) = 216.5 ± 4.1 M⁻¹).

FIGS. 38A-38B shows: 38A) the partial ¹H NMR spectra; and 38B) thebinding analysis curve for receptor 1 (0.1 mM) titrated with a combinedsolution of methyl α-D-glucoside (500 mM) and receptor 1 (0.1 mM), inD₂O buffered with 10 mM phosphate buffer solution (pH 7.4) at 298 K.Changes in chemical shift (Δδ ppm) of peak at 7.63 ppm (denoted with •)were plotted against increasing guest concentration (mM). The calculatedvalues for the Δδ are overlaid with the observed values, which areeffectively indicative of no binding taking place.

FIGS. 39A-39C shows for receptor 1 (0.06 mM) titrated withmethyl-α-D-glucoside (500 mM) in H₂O, in which: 39A) shows the blank ITCrun (addition of sugar into water); 39B) shows the actual run (sugarinto receptor 1); and 39C) shows the plotted change in enthalpy vs molarratio.

FIGS. 40A-40C shows the ITC binding results for receptor 1 (0.06 mM)titrated with methyl-α-D-glucoside (500 mM) in 10 mM phosphate buffersolution (pH 7.4), in which: 40A) shows the blank ITC run (addition ofsugar into water); 40B) shows the actual run (sugar into receptor 1) and40C) shows the plotted change in enthalpy vs molar ratio.

FIGS. 41A-41C shows the ITC results for receptor 1 (0.06 mM) titratedwith N-acetyl-D-glucosamine (498 mM) in H₂O, in which: 41A) shows theblank ITC run (addition of sugar into water); 41B) shows the actual run(sugar into receptor 1); and 41C) shows the plotted change in enthalpyvs molar ratio.

FIGS. 42A-42C shows the ITC binding results for receptor 1 (0.06 mM)titrated with N-acetyl-D-glucosamine (498 mM) in 10 mM phosphate buffersolution (pH 7.4), in which: 42A) shows the blank ITC run (addition ofsugar into water); 42B) shows the actual run (sugar into receptor 1) and42C) shows the plotted change in enthalpy vs molar ratio.

FIGS. 43A-43C shows for receptor 1 (0.06 mM) titrated with D-uracil (5mM) in 10 mM PBS buffer (pH 7.4), in which: 43A) shows the blank ITC run(addition of sugar into water); 43B) shows the actual run (sugar intoreceptor 1); and 43C) shows the plotted change in enthalpy vs molarratio.

FIGS. 44A-44C shows for receptor 1 (0.06 mM) titrated with uric acid(2.34 mM) in 10 mM PBS buffer (pH 7.4), in which: 44A) shows the blankITC run (addition of sugar into water); 44B) shows the actual run (sugarinto receptor 1); and 44C) shows the plotted change in enthalpy vs molarratio.

FIGS. 45A-45C shows the ITC results for receptor 1 (0.06 mM) titratedwith maltose (500 mM) in H₂O, in which: 45A) shows the blank ITC run(addition of sugar into water); 45B) shows the actual run (sugar intoreceptor 1); and 45C) shows the plotted change in enthalpy vs molarratio.

FIGS. 46A-46C shows the ITC binding results for receptor 1 (0.1 mM)titrated with D-Mannitol (500 mM) in 10 mM phosphate buffer solution (pH7.4) in which: 46A) shows the blank ITC run (addition of substrate intomedium); 46B) shows the actual run (substrate into receptor 1); 46C)shows the plotted change in enthalpy vs molar ratio.

FIGS. 47A-47C shows the ITC binding results for receptor 1 (0.06 mM)titrated with paracetamol (87 mM) in 10 mM phosphate buffer solution (pH7.4), in which: 47A) shows the blank ITC run (addition of substrate intowater); 47B) shows the actual run (substrate into receptor 1); 47C)shows the plotted change in enthalpy vs molar ratio.

FIGS. 48A-48C shows the ITC binding results for receptor 1 (0.06 mM)titrated with ascorbic acid (500 mM) in 10 mM phosphate buffer solution(pH 7.4), in which: 48A) shows the blank ITC run (addition of substrateinto water); 48B) shows the actual run (substrate into receptor 1); 48C)shows the plotted change in enthalpy vs molar ratio.

FIGS. 49A-49C shows the ITC binding results for receptor 1 (0.06 mM)titrated with L-fucose (500 mM) in 10 mM phosphate buffer solution (pH7.4), in which: 49A) shows the blank ITC run (addition of substrate intowater); 49B) shows the actual run (substrate into receptor 1); 49C)shows the plotted change in enthalpy vs molar ratio.

FIGS. 50A-50C shows the ITC binding results for receptor 1 (0.06 mM)titrated with L-phenylalanine (82 mM) in 10 mM phosphate buffer solution(pH 7.4), in which: 50A) shows the blank ITC run (addition of substrateinto medium); 50B) shows the actual run (substrate into receptor 1);50C) shows the plotted change in enthalpy vs molar ratio.

FIGS. 51A-51D shows the ITC binding results for receptor 1 (0.1 mM)titrated with myo inositol (5 mM) in 10 mM phosphate buffer solution (pH7.4) in which: 51A) shows the blank ITC run (addition of substrate intomedium); 51B) shows the actual run (substrate into receptor 1); 51C)shows the plotted change in enthalpy vs molar ratio; and 51D) shows thefit calculated using an Excel spreadsheet (K_(a) = 7563 ± 313 M⁻¹).

FIGS. 52A-52C shows the ITC binding results for receptor 1 (0.1 mM)titrated with Adenosine (500 mM) in 10 mM phosphate buffer solution (pH7.4) in which: 52A) shows the blank ITC run (addition of substrate intomedium); 52B) shows the actual run (substrate into receptor 1); 52C)shows the plotted change in enthalpy vs molar ratio.

FIGS. 53A-53C shows the ITC binding results for receptor 1 (0.1 mM)titrated with cytosine (20 mM) in 10 mM phosphate buffer solution (pH7.4) in which: 53A) shows the blank ITC run (addition of substrate intomedium); 53B) shows the actual run (substrate into receptor 1); 53C)shows the plotted change in enthalpy vs molar ratio.

FIGS. 54A-54C shows the ITC binding results for receptor 1 (0.06 mM)titrated with L-tryptophan (54 mM) in 10 mM phosphate buffer solution(pH 7.4), in which: 54A) shows the blank ITC run (addition of substrateinto medium);54 B) shows the actual run (substrate into receptor 1);54C) shows the plotted change in enthalpy vs molar ratio.

FIG. 55 shows the partial ¹H NMR ROESY spectrum of receptor 1 (2 mM)with D-glucose (5 mM, 2.5 equivalents) in D₂O. Chemical exchange peaks(black, annotated) link CH protons on β-D-glucose in free and boundstates. Chemical shifts for the glucose protons, with signal movementsdue to binding, are listed in the table. Signals for bound α-D-glucosewere not observed under these conditions.

FIG. 56 shows the structures of the substrates tested for affinity withreceptor 1.

FIGS. 57A-57B shows the partial ¹H NMR spectra (57A) and bindinganalysis curve (57B) for 90 (1 mM) titrated with a combined solution ofD-glucose (1 M) and 90 (1 mM), in D₂O with at pH 7.4 and 298 K. Changein chemical shifts (Δδ, ppm) denoted with • were plotted againstD-glucose concentration (mM). The calculated values for Δδ are overlaidwith the observed values, giving K_(a) = 5.1 ± 0.2 M-1 (3.6%).

FIGS. 58A-58B shows the Partial ¹H NMR spectra (58A) and bindinganalysis curve (58B) for 90 (0.25 mM) titrated with a combined solutionof D-cellobiose (250 mM) and 90 (0.25 mM), in D₂O with at pH 7.4 and 298K. Change in chemical shifts (Δδ, ppm) denoted with • were plottedagainst D-cellobiose concentration (mM). The calculated values for Δδare overlaid with the observed values, giving K_(a) = 46 ± 0.4 M-1(0.89%).

FIGS. 59A-59B shows the partial ¹H NMR spectra (59A) and bindinganalysis curve (59B) for 90 (0.2 mM) titrated with a combined solutionof D-cellotriose (15 mM) and 90 (0.2 mM), in D₂O with at pH 7.4 and 298K. Change in chemical shifts (Δδ, ppm) denoted with • were plottedagainst D-cellotriose concentration (mM). The calculated values for Δδare overlaid with the observed values, giving K_(a) = 949 ± 2.9 M-1(0.3%).

FIG. 60 shows the partial ¹H NMR spectra for 90 (0.2 mM) titrated with acombined solution of D-cellotetraose (15 mM) and 90 (0.2 mM), in D₂Owith at pH 7.4 and 298 K. Spectra imply binding with intermediate rateof exchange, thus no K_(a) was determinable.

FIG. 61 shows the partial ¹H NMR spectra for 90 (0.2 mM) titrated with acombined solution of D-cellopentaose (15 mM) and 90 (0.2 mM), in D₂Owith at pH 7.4 and 298 K. Spectra imply binding with intermediate rateof exchange, thus no K_(a) was determinable.

FIGS. 62A-62B shows the partial ¹H NMR spectra (62A) and bindinganalysis curve (62B) for 90 (0.2 mM) titrated with a combined solutionof D-maltose (500 mM) and 90 (0.2 mM), in D₂O with at pH 7.4 and 298 K.Change in chemical shifts (Δδ, ppm) denoted with • were plotted againstD-maltose concentration (mM). The calculated values for Δδ are overlaidwith the observed values, giving K_(a) = 15 ± 1.8 M-1 (11.8%).

FIGS. 63A-63B shows the partial ¹H NMR spectra (63A) and bindinganalysis curve (63B) for 90 (0.2 mM) titrated with a combined solutionof D-maltotriose (500 mM) and 90 (0.2 mM), in D₂O with at pH 7.4 and 298K. Change in chemical shifts (Δδ, ppm) denoted with • were plottedagainst D-maltotriose concentration (mM). The calculated values for Δδare overlaid with the observed values, giving K_(a) = 20 ± 0.7 M-1(3.3%).

FIGS. 64A-64D shows the ITC binding results for 90 (0.2 mM) titratedwith D-cellobiose (200 mM) in water at 298 K, in which: 64A) shows theblank run (addition of substrate into water); 64B) shows the titration(substrate into receptor); 64C) shows the plotted change in enthalpy vsmolar ratio; and 64D) shows the fit calculated using an Excelspreadsheet (K_(a) = 37.6 ± 2.5 M⁻¹).

FIGS. 65A-65D shows the ITC binding results for 90 (0.2 mM) titratedwith D-cellotriose (15 mM) in water at 298 K, in which: 65A) shows theblank run (addition of substrate into water); 65B) shows the titration(substrate into receptor); 65C) shows the plotted change in enthalpy vsmolar ratio; and 65D) shows the fit calculated using an Excelspreadsheet (K_(a) = 955 ± 11 M⁻¹).

FIGS. 66A-66B shows: 66A) the ITC titration of d-glucose (7.1 mM) in 10mM phosphate buffer into Receptor 4 (0.40 mM) in 10 mM. at 298 K; and66B) an enlarged image of the kcal mol⁻¹ of injectant vs molar ratiotrace. K_(a) calculated at 6490 M-1 +/- 72.6 M⁻¹.

FIGS. 67A-67B shows: 67A) the ITC titration of d-glucose (7.1 mM) in 10mM phosphate buffer into Receptor 5 (0.46 mM) in 10 mM. at 298 K; and67B) an enlarged image of the kcal mol⁻¹ of injectant vs molar ratiotrace. K_(a) calculated at 10400 M-1 +/- 132 M⁻¹.

FIG. 68 shows the ¹H NMR binding analysis curve generated following thetitration of a combined solution of β-D-glucose (10 mM) and Receptor 7(127 µM), in 10 mM PB, 140 mM NaCl, D₂O, into a solution of Receptor 7(127 µM) in 10 mM PB, 140 mM NaCl, D₂O. K_(a) calculated at 6886 M⁻¹ +/-190 M⁻¹.

FIGS. 69A-69B shows: 69A) the ITC titration of d-glucose (7.1 mM) in 10mM phosphate buffer into Receptor 8 (0.42 mM) in 10 mM. at 298 K; and69B) an enlarged image of the kcal mol⁻¹ of injectant vs molar ratiotrace. K_(a) calculated at 4210 M⁻¹ +/- 73 M⁻¹.

FIG. 70 shows the ¹H NMR binding analysis curve generated following thetitration of a combined solution of β-D-glucose (10 mM) and Recepor 9(210 µM), in 10 mM PB, 140 mM NaCl, D₂O, into a solution of Receptor 9(210 µM) in 10 mM PB, 140 mM NaCl, D₂O.

FIG. 71 shows the ¹H NMR binding analysis curve generated following thetitration of a combined solution of β-D-glucose (100 mM) and Recepor 10(250 µM), in 10 mM PB, 140 mM NaCl, D₂O, into a solution of Receptor 10(250 µM) in 10 mM PB, 140 mM NaCl, D₂O.

FIG. 72 shows the ¹H NMR binding analysis curve generated following thetitration of a combined solution of β-D-glucose (10 mM) and Recepor 13(265 µM), in 10 mM PB, 140 mM NaCl, D₂O, into a solution of Receptor 13(265 µM) in 10 mM PB, 140 mM NaCl, D₂O.

FIG. 73 shows the the partial ¹H NMR spectra for Receptor 11 (50 µM) inD₂O (pH 7.4, 10 mM PBsoln) titrated with D-glucose (10 mM) with addedReceptor 11 (50 µM) and 10 mM PBsoln. In making the assumption ofreceptor saturation at ~1 mM, half saturation would be at 0.5 mM.Therefore 1/0.5 mM = K_(a) ~ 2000 M⁻¹.

FIGS. 74A-74B shows: 74A) the circular dichroism (CD) spectra; and 74B)the binding analysis curve generated following the titration ofD-glucose (10 mM) with added Receptor 11 (70 µM) and 10 mM PBsoln to asolution of Receptor 11 (70 µM) in water (pH 7.4 with 10 mM PBsoln).

FIGS. 75A-75B shows: 75A) the ITC titration of d-glucose (7.73 mM) in 10mM phosphate buffer into Receptor 13 (0.13 mM) in 10 mM at 298 K; and75B) an enlarged image of the kcal mol⁻¹ of injectant vs molar ratiotrace. K_(a) calculated at 1310 M⁻¹ +/- 33 M⁻¹.

FIGS. 76A-76B shows: 76A) the ITC titration of d-glucose (7.10 mM) in 10mM phosphate buffer into Receptor 3 (0.29 mM) in 10 mM. at 298 K; and76B) an enlarged image of the kcal mol⁻¹ of injectant vs molar ratiotrace. K_(a) calculated at 5760 M⁻¹ +/- 269 M⁻¹.

FIG. 77 shows the ¹H NMR binding analysis curve generated following thetitration of a combined solution of β-D-glucose (3.24 M) and Recepor 2(265 µM) in D₂O, into a solution of Receptor 12 (223 µM) in D₂O at 298K.

DETAILED DESCRIPTION OF THE INVENTION Definitions

Unless otherwise stated, the following terms used in the specificationand claims have the following meanings set out below.

Throughout the description and claims of this specification, the words“comprise” and “contain” and variations of them mean “including but notlimited to”, and they are not intended to (and do not) exclude othermoieties, additives, components, integers or steps. Throughout thedescription and claims of this specification, the singular encompassesthe plural unless the context otherwise requires. In particular, wherethe indefinite article is used, the specification is to be understood ascontemplating plurality as well as singularity, unless the contextrequires otherwise.

Features, integers, characteristics, compounds, chemical moieties orgroups described in conjunction with a particular aspect, embodiment orexamples of the invention are to be understood to be applicable to anyother aspect, embodiment or example described herein unless incompatibletherewith. All of the features disclosed in this specification(including any accompanying claims, abstract and drawings), and/or allof the steps of any method or process so disclosed, may be combined inany combination, except combinations where at least some of suchfeatures and/or steps are mutually exclusive. The invention is notrestricted to the details of any foregoing embodiments. The inventionextends to any novel one, or any novel combination, of the featuresdisclosed in this specification (including any accompanying claims,abstract and drawings), or to any novel one, or any novel combination,of the steps of any method or process so disclosed.

In this specification the term “alkyl” includes both straight andbranched chain alkyl groups. References to individual alkyl groups suchas “propyl” are specific for the straight chain version only andreferences to individual branched chain alkyl groups such as “isopropyl”are specific for the branched chain version only. For example,“(1-6C)alkyl” includes (1-4C)alkyl, (1-3C)alkyl, propyl, isopropyl andt-butyl. A similar convention applies to other radicals, for example“phenyl(1-6C)alkyl” includes phenyl(1-4C)alkyl, benzyl, 1-phenylethyland 2-phenylethyl.

The term “alkenyl” will be understood to include both straight andbranched hydrocarbon groups comprising one or more carbon-carbon doublebonds. Reference to, for example, “(2-6C)alkenyl” will be understood torefer to alkene groups containing from 3 to 6 carbon atoms and mayincludes, for example, hexenyl, pentenyl, butenyl, propenyl andethylenyl.

The term “alkynyl” will be understood to include both straight andbranched hydrocarbon groups comprising one or more carbon-carbon triplebonds. Again, reference to “(2-6C)alkynyl” groups will be understood torefer to alkyne groups containing from 3 to 6 carbon atoms and mayincludes, for example hexynyl, pentynyl, butynyl, propynyl andacetylenyl.

The term “(m-nC)” or “(m-nC) group” used alone or as a prefix, refers toany group having m to n carbon atoms.

An “alkylene,” “alkenylene,” or “alkynylene” group is an alkyl, alkenyl,or alkynyl group that is positioned between and serves to connect twoother chemical groups. Thus, “(1-6C)alkylene” means a linear saturateddivalent hydrocarbon radical of one to six carbon atoms or a branchedsaturated divalent hydrocarbon radical of three to six carbon atoms, forexample, methylene, ethylene, propylene, 2-methylpropylene, pentylene,and the like.

“(3-8C)cycloalkyl” means a hydrocarbon ring containing from 3 to 8carbon atoms, for example, cyclopropyl, cyclobutyl, cyclopentyl,cyclohexyl, cycloheptyl or bicyclo[2.2.1]heptyl.

“(3-8C)cycloalkenyl” means a hydrocarbon ring containing from 3 to 8carbon atoms and at least one double bond, for example, cyclobutenyl,cyclopentenyl, cyclohexenyl or cycloheptenyl, such as 3-cyclohexen-1-yl,or cyclooctenyl.

The term “heterocyclyl”, “heterocyclic” or “heterocycle” means anon-aromatic saturated or partially saturated monocyclic, fused,bridged, or spiro bicyclic heterocyclic ring system(s). Monocyclicheterocyclic rings contain from about 3 to 12 (suitably from 3 to 7)ring atoms, with from 1 to 5 (suitably 1, 2 or 3) heteroatoms selectedfrom nitrogen, oxygen or sulfur in the ring. Bicyclic heterocyclescontain from 7 to 17 member atoms, suitably 7 to 12 member atoms, in thering. Bicyclic heterocyclic(s) rings may be fused, spiro, or bridgedring systems. Examples of heterocyclic groups include cyclic ethers suchas oxiranyl, oxetanyl, tetrahydrofuranyl, dioxanyl, and substitutedcyclic ethers. Heterocycles containing nitrogen include, for example,azetidinyl, pyrrolidinyl, piperidinyl, piperazinyl, tetrahydrotriazinyl,tetrahydropyrazolyl, and the like. Typical sulfur containingheterocycles include tetrahydrothienyl, dihydro-1,3-dithiol,tetrahydro-2H-thiopyran, and hexahydrothiepine. Other heterocyclesinclude dihydro-oxathiolyl, tetrahydro-oxazolyl, tetrahydro-oxadiazolyl,tetrahydrodioxazolyl, tetrahydro-oxathiazolyl, hexahydrotriazinyl,tetrahydro-oxazinyl, morpholinyl, thiomorpholinyl,tetrahydropyrimidinyl, dioxolinyl, octahydrobenzofuranyl,octahydrobenzimidazolyl, and octahydrobenzothiazolyl. For heterocyclescontaining sulfur, the oxidized sulfur heterocycles containing SO or SO₂groups are also included. Examples include the sulfoxide and sulfoneforms of tetrahydrothienyl and thiomorpholinyl such as tetrahydrothiene1,1-dioxide and thiomorpholinyl 1,1-dioxide. A suitable value for aheterocyclyl group which bears 1 or 2 oxo (=O) or thioxo (=S)substituents is, for example, 2-oxopyrrolidinyl, 2-thioxopyrrolidinyl,2-oxoimidazolidinyl, 2-thioxoimidazolidinyl, 2-oxopiperidinyl,2,5-dioxopyrrolidinyl, 2,5-dioxoimidazolidinyl or 2,6-dioxopiperidinyl.Particular heterocyclyl groups are saturated monocyclic 3 to 7 memberedheterocyclyls containing 1, 2 or 3 heteroatoms selected from nitrogen,oxygen or sulfur, for example azetidinyl, tetrahydrofuranyl,tetrahydropyranyl, pyrrolidinyl, morpholinyl, tetrahydrothienyl,tetrahydrothienyl 1,1-dioxide, thiomorpholinyl, thiomorpholinyl1,1-dioxide, piperidinyl, homopiperidinyl, piperazinyl orhomopiperazinyl. As the skilled person would appreciate, any heterocyclemay be linked to another group via any suitable atom, such as via acarbon or nitrogen atom. However, reference herein to piperidino ormorpholino refers to a piperidin-1-yl or morpholin-4-yl ring that islinked via the ring nitrogen.

By “bridged ring systems” is meant ring systems in which two rings sharemore than two atoms, see for example Advanced Organic Chemistry, byJerry March, 4^(th) Edition, Wiley Interscience, pages 131-133, 1992.Examples of bridged heterocyclyl ring systems include,aza-bicyclo[2.2.1]heptane, 2-oxa-5-azabicyclo[2.2.1]heptane,aza-bicyclo[2.2.2]octane, aza-bicyclo[3.2.1]octane and quinuclidine.

By “spiro bi-cyclic ring systems” we mean that the two ring systemsshare one common spiro carbon atom, i.e. the heterocyclic ring is linkedto a further carbocyclic or heterocyclic ring through a single commonspiro carbon atom. Examples of spiro ring systems include6-azaspiro[3.4]octane, 2-oxa-6-azaspiro[3.4]octane,2-azaspiro[3.3]heptanes, 2-oxa-6-azaspiro[3.3]heptanes,7-oxa-2-azaspiro[3.5]nonane, 6-oxa-2-azaspiro[3.4]octane,2-oxa-7-azaspiro[3.5]nonane and 2-oxa-6-azaspiro[3.5]nonane.

“Heterocyclyl(1-6C)alkyl” means a heterocyclyl group covalently attachedto a (1-6C)alkylene group, both of which are defined herein.

The term “heteroaryl” or “heteroaromatic” means an aromatic mono-, bi-,or polycyclic ring incorporating one or more (for example 1-4,particularly 1, 2 or 3) heteroatoms selected from nitrogen, oxygen orsulfur. The term heteroaryl includes both monovalent species anddivalent species. Examples of heteroaryl groups are monocyclic andbicyclic groups containing from five to twelve ring members, and moreusually from five to ten ring members. The heteroaryl group can be, forexample, a 5- or 6-membered monocyclic ring or a 9- or 10-memberedbicyclic ring, for example a bicyclic structure formed from fused fiveand six membered rings or two fused six membered rings. Each ring maycontain up to about four heteroatoms typically selected from nitrogen,sulfur and oxygen. Typically, the heteroaryl ring will contain up to 3heteroatoms, more usually up to 2, for example a single heteroatom. Inone embodiment, the heteroaryl ring contains at least one ring nitrogenatom. The nitrogen atoms in the heteroaryl rings can be basic, as in thecase of an imidazole or pyridine, or essentially non-basic as in thecase of an indole or pyrrole nitrogen. In general, the number of basicnitrogen atoms present in the heteroaryl group, including any aminogroup substituents of the ring, will be less than five.

Examples of heteroaryl include furyl, pyrrolyl, thienyl, oxazolyl,isoxazolyl, imidazolyl, pyrazolyl, thiazolyl, isothiazolyl, oxadiazolyl,thiadiazolyl, triazolyl, tetrazolyl, pyridyl, pyridazinyl, pyrimidinyl,pyrazinyl, 1,3,5-triazenyl, benzofuranyl, indolyl, isoindolyl,benzothienyl, benzoxazolyl, benzimidazolyl, benzothiazolyl,benzothiazolyl, indazolyl, purinyl, benzofurazanyl, quinolyl,isoquinolyl, quinazolinyl, quinoxalinyl, cinnolinyl, pteridinyl,naphthyridinyl, carbazolyl, phenazinyl, benzisoquinolinyl,pyridopyrazinyl, thieno[2,3-b]furanyl, 2H-furo[3,2-b]-pyranyl,5H-pyrido[2,3-d]-o-oxazinyl, 1H-pyrazolo[4,3-d]-oxazolyl,4H-imidazo[4,5-d]thiazolyl, pyrazino[2,3-d]pyridazinyl,imidazo[2,1-b]thiazolyl, imidazo[1,2-b][1,2,4]triazinyl. “Heteroaryl”also covers partially aromatic bi- or polycyclic ring systems wherein atleast one ring is an aromatic ring and one or more of the other ring(s)is a non-aromatic, saturated or partially saturated ring, provided atleast one ring contains one or more heteroatoms selected from nitrogen,oxygen or sulfur. Examples of partially aromatic heteroaryl groupsinclude for example, tetrahydroisoquinolinyl, tetrahydroquinolinyl,2-oxo-1,2,3,4-tetrahydroquinolinyl, dihydrobenzthienyl,dihydrobenzfuranyl, 2,3-dihydro-benzo[1,4]dioxinyl, benzo[1,3]dioxolyl,2,2-dioxo-1,3-dihydro-2-benzothienyl, 4,5,6,7-tetrahydrobenzofuranyl,indolinyl, 1,2,3,4-tetrahydro-1,8-naphthyridinyl,1,2,3,4-tetrahydropyrido[2,3-b]pyrazinyl and3,4-dihydro-2H-pyrido[3,2-b][1,4]oxazinyl.

Examples of five membered heteroaryl groups include but are not limitedto pyrrolyl, furanyl, thienyl, imidazolyl, furazanyl, oxazolyl,oxadiazolyl, oxatriazolyl, isoxazolyl, thiazolyl, isothiazolyl,pyrazolyl, triazolyl and tetrazolyl groups.

Examples of six membered heteroaryl groups include but are not limitedto pyridyl, pyrazinyl, pyridazinyl, pyrimidinyl and triazinyl.

A bicyclic heteroaryl group may be, for example, a group selected from:

-   a benzene ring fused to a 5- or 6-membered ring containing 1, 2 or 3    ring heteroatoms;-   a pyridine ring fused to a 5- or 6-membered ring containing 1, 2 or    3 ring heteroatoms;-   a pyrimidine ring fused to a 5- or 6-membered ring containing 1 or 2    ring heteroatoms;-   a pyrrole ring fused to a 5- or 6-membered ring containing 1, 2 or 3    ring heteroatoms;-   a pyrazole ring fused to a 5- or 6-membered ring containing 1 or 2    ring heteroatoms;-   a pyrazine ring fused to a 5- or 6-membered ring containing 1 or 2    ring heteroatoms;-   an imidazole ring fused to a 5- or 6-membered ring containing 1 or 2    ring heteroatoms;-   an oxazole ring fused to a 5- or 6-membered ring containing 1 or 2    ring heteroatoms;-   an isoxazole ring fused to a 5- or 6-membered ring containing 1 or 2    ring heteroatoms;-   a thiazole ring fused to a 5- or 6-membered ring containing 1 or 2    ring heteroatoms;-   an isothiazole ring fused to a 5- or 6-membered ring containing 1 or    2 ring heteroatoms;-   a thiophene ring fused to a 5- or 6-membered ring containing 1, 2 or    3 ring heteroatoms;-   a furan ring fused to a 5- or 6-membered ring containing 1, 2 or 3    ring heteroatoms;-   a cyclohexyl ring fused to a 5- or 6-membered heteroaromatic ring    containing 1, 2 or 3 ring heteroatoms; and-   a cyclopentyl ring fused to a 5- or 6-membered heteroaromatic ring    containing 1, 2 or 3 ring heteroatoms.

Particular examples of bicyclic heteroaryl groups containing asix-membered ring fused to a five-membered ring include but are notlimited to benzfuranyl, benzthiophenyl, benzimidazolyl, benzoxazolyl,benzisoxazolyl, benzthiazolyl, benzisothiazolyl, isobenzofuranyl,indolyl, isoindolyl, indolizinyl, indolinyl, isoindolinyl, purinyl(e.g., adeninyl, guaninyl), indazolyl, benzodioxolyl andpyrazolopyridinyl groups.

Particular examples of bicyclic heteroaryl groups containing two fusedsix membered rings include but are not limited to quinolinyl,isoquinolinyl, chromanyl, thiochromanyl, chromenyl, isochromenyl,chromanyl, isochromanyl, benzodioxanyl, quinolizinyl, benzoxazinyl,benzodiazinyl, pyridopyridinyl, quinoxalinyl, quinazolinyl, cinnolinyl,phthalazinyl, naphthyridinyl and pteridinyl groups.

The term “aryl” means a cyclic or polycyclic aromatic ring having from 5to 12 carbon atoms. The term aryl includes both monovalent species anddivalent species. Examples of aryl groups include, but are not limitedto, phenyl, biphenyl, naphthyl, anthraceneyl and the like. In particularembodiment, an aryl is phenyl.

The term “halo” refers to any suitable halogen and may be selected fromfluoro, chloro, bromo and iodo groups. Suitably, the term halo refers tofluoro, chloro or bromo groups, and most suitably, chloro groups.

The term “optionally substituted” refers to either groups, structures,or molecules that are substituted and those that are not substituted.The term “wherein a/any CH, CH₂, CH₃ group or heteroatom (i.e. NH)within a R¹ group is optionally substituted” suitably means that (any)one of the hydrogen radicals of the R¹ group is substituted by arelevant stipulated group.

The term “hydrophilic substituent group” will be understood to refer toa substituent group which has an affinity for water and tends tosolvation. Accordingly, the term “hydrophilic substituent group” may beunderstood to encompass any substituent group which facilitates watersolubility to the compounds of the present invention.

The term “hydrophilic polymer” will be understood to refer to anyoligomer, polymer and/or copolymer comprising at least 3 repeat units(suitably at least 10 repeat units), wherein one or more of said repeatunits (monomers) comprise a polar functional group with an affinity forwater. The term “hydrophilic polymer” will be understood to encompasslinear, branched and hyperbranched polymers. Suitably, the hydrophilicpolymer is selected from a polycarboxylic acid, polycarboxylate,polyhydroxy, polyester, polyether, polyamine, polyamide, polyphosphateor polyoxyalkylene. More suitably, a polycarboxylic acid,polycarboxylate, polyhydroxy or polyether. Yet more suitably, thehydrophilic polymer is polyethylene glycol, polyvinylalcohol,polyacrylate, polyacrylamide or polyvinyl pyrrolidine. Most suitably,the hydrophilic polymer is polyethylene glycol or polyacrylamide.

The term “hydrophilic dendritric group” will be understood to refer toany dendrimer, dendron or branched molecule comprising one or more polarfunctional group with an affinity for water. That is, the term“hydrophilic dendritric group” will be understood to encompass anydendrimer, dendron or branched molecule which facilitates the watersolublity of the compounds of the present invention. Moreover, the term“dendrimer” is a term of the art and will readily be understood to referto a tree-like molecular architecture with a core or focal point,interior layers (otherwise known as “generations”) which consist ofrepeating units of one or more building units attached to the core orfocal point, and an exterior (outermost) layer of building unitscomprising terminal functional groups on the terminus of the dendrimerstructure.

Where optional substituents are chosen from “one or more” groups it isto be understood that this definition includes all substituents beingchosen from one of the specified groups or the substituents being chosenfrom two or more of the specified groups.

The phrase “compound of the invention” means those compounds which aredisclosed herein, both generically and specifically.

Compounds of the Invention

According to one aspect of the present invention, there is provided acompound of Formula (I), or a salt, hydrate or solvate thereof, as shownbelow:

wherein:

-   bonds b₁ and b₂ are independently selected from a single bond or    double bond;

-   R_(1a), R_(1b), R_(2a) and R_(2b) are independently selected from    hydrogen, carbonyl, (1-8C)alkyl, (3-10C)cycloalkyl, aryl, heteroaryl    and heterocyclyl, each of which, other than hydrogen and carbonyl,    is optionally substituted by one or more substituent groups selected    from (1-4C)alkyl, halo, (1-4C)haloalkyl, (1-4C)haloalkoxy,    (1-4C)alkoxy, (1-4C)alkylamino, amino, cyano, hydroxyl, carboxy,    carbamoyl, sulfamoyl, mercapto and a hydrophilic substituent group;    or

-   R_(1a) and R_(1b) are linked so as to form a group of the formula:

-   

-   and/or R_(2a) and R_(2b) are linked so as to form a group of the    formula:

-   

-   wherein:    -   

    -   denotes the point of attachment;

    -   bonds b₁ and b₂ are as described above;

    -   Rings A and B are independently selected from aryl, heteroaryl,        heterocyclyl, cycloalkyl and cycloalkenyl;

    -   R₁ and R₂ are independently selected from (1-6C)alkyl, halo,        (1-4C)haloalkyl, (1-4C)haloalkoxy, (1-6C)alkoxy,        (1-4C)alkylamino, amino, cyano, hydroxyl, carboxy, carbamoyl,        sulfamoyl and mercapto;

    -   a and b are integers independently selected from 0 to 2;

    -   m and n are integers independently selected from 0 to 2;

    -   Z₁ and Z₂ are independently selected from a hydrophilic        substituent group;

-   C and D are independently selected from aryl, heteroaryl,    heterocyclyl, cycloalkyl, cycloalkenyl and a group of the formula:

-   

-   wherein:    -   s, t and v are integers independently selected from 1 or 2;

    -   

    -   denotes the point of attachment;

-   R₃ and R₄ are independently selected from halo, (1-4C)alkyl,    (1-4C)alkoxy, amino, nitro, (1-4C)alkylamino, (1-4C)dialkylamino,    (1-4C)haloalkyl, (1-4C)haloalkoxy, cyano, (2-4C)alkenyl,    (2-4C)alkynyl and a group of the formula:

-   

-   wherein:    -   L¹ is absent or a (1-5C)alkylene optionally substituted by one        or more substituents selected from (1-2C)alkyl and oxo;

    -   Y¹ is absent or selected from a one of the following groups; O,        S, SO, SO₂, N(R_(a)), C(O), C(O)O, OC(O), C(O)N(R_(a)),        N(R_(a))C(O), N(R_(b))C(O)N(R_(a)), N(R_(a))C(O)O,        OC(O)N(R_(a)), S(O)₂N(R_(a)), and N(R_(a))SO₂, wherein R_(a) and        R_(b) are each independently selected from hydrogen and        (1-4C)alkyl; and

    -   Q¹ is hydrogen, (1-8C)alkyl, (2-6C)alkenyl, (2-6C)alkynyl, aryl,        (3-10C)cycloalkyl, (3-10C)cycloalkenyl, heteroaryl and        heterocyclyl; wherein

    -   Q¹ is optionally further substituted by one or more substituent        groups independently selected from (1-4C)alkyl, halo,        (1-4C)haloalkyl, (1-4C)haloalkoxy, amino, (1-4C)aminoalkyl,        cyano, hydroxy, carboxy, carbamoyl, sulfamoyl, mercapto, ureido,        oxy, NR_(c)R_(d), OR_(c), C(O)R_(d), C(O)OR_(c), OC(O)R_(c),        C(O)N(R_(d))R_(c), N(R_(d))C(O)R_(c), S(O)_(y)R_(c) (where y is        0, 1 or 2), SO₂N(R_(d))R_(c), N(R_(d))SO₂R_(c),        Si(R_(e))(R_(d))R_(c) and (CH₂)_(z)NR_(d)R_(c) (where z is 1, 2        or 3); wherein R_(c), R_(d) and R_(e) are each independently        selected from hydrogen, (1-6C)alkyl and (3-6C)cycloalkyl; and        R_(c) and R_(d) can be linked such that, together with the        nitrogen atom to which they are attached, they form a 4-7        membered heterocyclic ring which is optionally substituted by        one or more substituents selected from (1-4C)alkyl, halo,        (1-4C)haloalkyl, (1-4C)haloalkoxy, (1-4C)alkoxy,        (1-4C)alkylamino, amino, cyano or hydroxyl; or

    -   two R₃ and/or two R₄ groups taken together may form a group of        the formula:

    -   

    -   wherein:        -   R_(x) is selected from hydrogen and (1-6C)alkyl optionally            substituted by one or more substituent groups selected from            halo, (1-4C)haloalkyl, (1-4C)haloalkoxy, cyano, hydroxy,            sulfamoyl, mercapto, ureido, NR_(f)R_(g), OR_(f), C(O)R_(f),            C(O)OR_(f), OC(O)R_(f), C(O)N(R_(g))R_(f) and            N(R_(g))C(O)R_(f), wherein R_(f) and R_(g) are selected from            hydrogen and (1-4C)alkyl; and        -   dashed lines represent the points of attachment to C and/or            D;

-   W₁, W₂, W₃ and W₄ are independently selected from CR_(h)R_(i),    wherein R_(h) and R_(i) are selected from hydrogen and (1-2C)alkyl;

-   X₁, X₂, X₃ and X₄ are independently selected from a group of the    formula:

-   

-   wherein:    -   

    -   denotes the point of attachment;

    -   W_(x) is selected from O or NH; and

    -   Q is selected from O, S and NR_(j), wherein R_(j) is selected        from hydrogen, (1-4C)alkyl, aryl, heteroaryl and sulfonyl;

-   Z₃ and Z₄ are independently selected from a hydrophilic substituent    group;

-   L is absent or a linker, which optionally bears a hydrophilic    substituent group Z₅;

-   c and d are integers independently selected from 0 to 4;

-   and p are integers independently selected from 0 t2; and wherein:    -   i) the compound of Formula I is optionally attached to a        displaceable reporter molecule via one or more of the        substituent groups associated with R₁, R₂, R₃, R₄, Z₁, Z₂, Z₃,        Z₄ and/or Z₅; and/or

    -   ii) the compound of Formula I is optionally attached to a        substituent group of Formula A1 shown below at a position        associated with one or more of the substituent groups R_(1a),        R_(1b), R_(2a), R_(2b), R₁, R₂, R₃, R₄, Z₁, Z₂, Z₃, Z₄ and/or        Z₅:

    -   

    -   wherein:        -   X_(2a) is absent or selected from O, S, SO, SO₂, N(R^(x2)),            C(O), C(O)O, OC(O), C(O)N(R^(x2)), N(R^(x2))C(O),            N(R^(x2))C(O)N(R^(x3)), N(R^(x2))C(O)O, OC(O)N(R^(x2)),            S(O)₂N(R^(x2)) and N(R^(x2))SO₂, wherein R^(x2) and R^(x3)            are each independently selected from hydrogen and            (1-4C)alkyl;

        -   L_(2a) is absent or selected from (1-20C)alkylene,            (1-20C)alkylene oxide, (1-20C)alkenyl and (1-20C)alkynyl,            each of which being optionally substituted by one or more            substituents selected from (1-2C)alkyl, aryl and oxo; and

        -   Z_(2a) is selected from carboxy, carbamoyl, sulphamoyl,            mercapto, amino, azido, (1-4C)alkenyl, (1-4C)alkynyl,            NR^(xc)R^(xd), OR^(xc), ONR^(xc)R^(xd), C(O)X_(a),            C(Q^(z))OR^(xf), N=C=O, NR^(xc)C(O)CH₂X_(b),            C(O)N(R^(xe))NR^(Xc)R^(Xd), S(O)_(y)X_(a) (where y is 0, 1            or 2), SO₂N(R^(xe))NR^(xc)R^(xd), Si(R^(xg))(R^(xh))R^(xi),            S-S-X_(c) an amino acid and

        -   

        -   wherein:            -   X_(a) is a leaving group (e.g. halo or CF₃);            -   X_(b) is a halo (e.g. iodo);            -   X_(c) is an aryl or heteroaryl, optionally substituted                with one or more substituents selected from halo, cyano                and nitro;            -   R^(xc), R^(xd) and R^(xe) are each independently                selected from hydrogen and (1-6C)alkyl;            -   R^(xf) is selected from hydrogen or (1-6C)alkyl, or                R^(xf) is a substituent group that renders C(O)OR^(xf),                when taken as a whole, to be an activated ester (e.g a                hydroxysuccinimide ester, a hydroxy-3-sulfo-succinimide                ester or a pentafluorophenyl ester);            -   Q^(z) is selected from O or ⁺NR^(Q1)R^(Q2), where R^(Q1)                and R^(Q2) are independently selected from hydrogen and                methyl; and R^(xg), R^(xh) and R^(xi) are each                independently selected from (1-4C)alkyl, hydroxy, halo                and (1-4C)alkoxy;

with the proviso that the compound of Formula I comprises at least onehydrophilic substituent group (e.g. Z₁, Z₂, Z₃, Z₄ or Z₅).

To their surprise, the inventors have advantageously discovered that thecompounds of the present invention display an exceptionally highaffinity towards certain target saccharides (e.g. glucose) in aqueousmedia. Furthermore, it has also been discovered that the compounds ofthe present invention display unprecedented levels of selectivitytowards certain target saccharides (e.g. glucose) over otherstructurally similar saccharides (e.g. mannose).

In addition to the remarkable affinities and selectivities displayed bythe compounds of the present invention, the non-covalent interactionsbetween the compounds of the present invention and the targetsaccharide, as opposed to covalent interactions, allow the compounds ofthe present invention to reversibly associate with certain targetsaccharides. Noting the aforementioned difficulties in the artassociated with the development of efficient and selective saccharidereceptor molecules, particularly in biologically relevant aqueous media,the compounds of the present invention clearly represent a unique andhighly useful class of compounds for use in saccharide sensingapplications.

Without wishing to be bound by theory, it is believed that one guidingprinciple in the design of the compounds of the present invention iscomplementarity. Suitably, both polar and apolar groups of the compoundsof the present invention are positioned to make favourable contacts withthe target saccharide. For instance, the substituent groups C and D arecapable of making hydrophobic/CH-π contacts with axial CH groups of thesaccharide, while the spacer groups (e.g. the bis-urea based motifs) arecapable of forming hydrogen bond interactions to —O— and —OH units ofthe saccharide. Furthermore, the bis-urea based spacer groups of thecompounds of the present invention also help to maintain a well-defined‘cavity’, holding substituent groups C and D the correct distance apartfor successful saccharide binding interactions to occur (i.e. between 8and 10 Ǻ, suitably, approximately 9 Ǻ). For illustration purposes only,FIG. 1 shows both schematic illustations of the key interactions madebetween the target saccharide and the compounds of the present invention(FIGS. 1 a and 1 b ), as well as molecular models of a ground stateconformation of one particular compound of the present invention withglucose (FIGS. 1 c and 1 d ). In FIG. 1 c , ten intermolecular NH···Ohydrogen bonds (with a distance of between 1.9 and 2.2 Å) can be seen,and FIG. 1 d further depicts the close CH-π contacts made between thesaccharide and compound of the present invention.

The compounds of the present invention also benefit from high levels ofwater solublity, thereby allowing them to readily dissolve in, and thusbe compatible for use in, the various aqueous media in which targetsaccharides commonly exist (i.e. in the bloodstream or in fermentationmedia). Thus, in an embodiment, the compounds of the present inventionare water soluble. Suitably, the compounds of the present invention havewater solubility of at least about 1 µM. More suitably, the compounds ofthe present invention have water solubility of at least about 100 µM.Yet more suitably, the compounds of the present invention have watersolubility of at least about 500 µM. Even more suitably, the compoundsof the present invention have water solubility of at least about 1 mM.Most suitably, the compounds of the present invention have watersolubility of at least about 2 mM.

In another embodiment, the compounds of the present invention have watersolubility of between 1 µM and 50 mM. Suitably, the compounds of thepresent invention have water solubility of between 1 µM and 20 mM. Moresuitably, the compounds of the present invention have water solubilityof between 100 µM and 20 mM. Most suitably, the compounds of the presentinvention have water solubility of between 100 µM and 10 mM.

In a further embodiment, there is provided a compound of Formula (I), ora salt, hydrate or solvate thereof, as shown below:

wherein:

-   bonds b₁ and b₂ are independently selected from a single bond or    double bond;

-   R_(1a), R_(1b), R_(2a) and R_(2b) are independently selected from    hydrogen, carbonyl, (1-8C)alkyl, (3-10C)cycloalkyl, aryl, heteroaryl    and heterocyclyl, each of which, other than hydrogen and carbonyl,    is optionally substituted by one or more substituent groups selected    from (1-4C)alkyl, halo, (1-4C)haloalkyl, (1-4C)haloalkoxy,    (1-4C)alkoxy, (1-4C)alkylamino, amino, cyano, hydroxyl, carboxy,    carbamoyl, sulfamoyl, mercapto and a hydrophilic substituent group;    or

-   R_(1a) and R_(1b) are linked so as to form a group of the formula:

-   

-   and/or R_(2a) and R_(2b) are linked so as to form a group of the    formula:

-   

-   wherein:    -   

    -   denotes the point of attachment;

    -   bonds b₁ and b₂ are as described above;

    -   Rings A and B are independently selected from aryl, heteroaryl,        heterocyclyl, cycloalkyl and cycloalkenyl;

    -   R₁ and R₂ are independently selected from (1-6C)alkyl, halo,        (1-4C)haloalkyl, (1-4C)haloalkoxy, (1-6C)alkoxy,        (1-4C)alkylamino, amino, cyano, hydroxyl, carboxy, carbamoyl,        sulfamoyl and mercapto;

    -   a and b are integers independently selected from 0 to 2;

    -   m and n are integers independently selected from 0 to 2;

    -   Z₁ and Z₂ are independently selected from a hydrophilic        substituent group;

-   C and D are independently selected from aryl, heteroaryl,    heterocyclyl, cycloalkyl, cycloalkenyl and a group of the formula:

-   

-   wherein:    -   s, t and v are integers independently selected from 1 or 2;

    -   

    -   denotes the point of attachment;

-   R₃ and R₄ are independently selected from halo, (1-4C)alkyl,    (1-4C)alkoxy, amino, nitro, (1-4C)alkylamino, (1-4C)dialkylamino,    (1-4C)haloalkyl, (1-4C)haloalkoxy, cyano, (2-4C)alkenyl,    (2-4C)alkynyl and a group of the formula:

-   

-   wherein:    -   L¹ is absent or a (1-5C)alkylene optionally substituted by one        or more substituents selected from (1-2C)alkyl and oxo;

    -   Y¹ is absent or selected from a one of the following groups; O,        S, SO, SO₂, N(R_(a)), C(O), C(O)O, OC(O), C(O)N(R_(a)),        N(R_(a))C(O), N(R_(b))C(O)N(R_(a)), N(R_(a))C(O)O,        OC(O)N(R_(a)), S(O)₂N(R_(a)), and N(R_(a))SO₂, wherein R_(a) and        R_(b) are each independently selected from hydrogen and        (1-4C)alkyl; and

    -   Q¹ is hydrogen, (1-8C)alkyl, (2-6C)alkenyl, (2-6C)alkynyl, aryl,        (3-10C)cycloalkyl, (3-10C)cycloalkenyl, heteroaryl and        heterocyclyl; wherein

    -   Q¹ is optionally further substituted by one or more substituent        groups independently selected from (1-4C)alkyl, halo,        (1-4C)haloalkyl, (1-4C)haloalkoxy, amino, (1-4C)aminoalkyl,        cyano, hydroxy, carboxy, carbamoyl, sulfamoyl, mercapto, ureido,        oxy, NR_(c)R_(d), OR_(c), C(O)R_(d), C(O)OR_(c), OC(O)R_(c),        C(O)N(R_(d))R_(c), N(R_(d))C(O)R_(c), S(O)_(y)R_(c) (where y is        0, 1 or 2), SO₂N(R_(d))R_(c), N(R_(d))SO₂R_(c),        Si(R_(e))(R_(d))R_(c) and (CH₂)_(z)NR_(d)R_(c) (where z is 1, 2        or 3); wherein R_(c), R_(d) and R_(e) are each independently        selected from hydrogen, (1-6C)alkyl and (3-6C)cycloalkyl; and        R_(c) and R_(d) can be linked such that, together with the        nitrogen atom to which they are attached, they form a 4-7        membered heterocyclic ring which is optionally substituted by        one or more substituents selected from (1-4C)alkyl, halo,        (1-4C)haloalkyl, (1-4C)haloalkoxy, (1-4C)alkoxy,        (1-4C)alkylamino, amino, cyano or hydroxyl; or

    -   two R₃ and/or two R₄ groups taken together form a group of the        formula:

    -   

    -   wherein:        -   R_(x) is selected from hydrogen and (1-6C)alkyl optionally            substituted by one or more substituent groups selected from            halo, (1-4C)haloalkyl, (1-4C)haloalkoxy, cyano, hydroxy,            sulfamoyl, mercapto, ureido, NR_(f)R_(g), OR_(f), C(O)R_(f),            C(O)OR_(f), OC(O)R_(f), C(O)N(R_(g))R_(f) and            N(R_(g))C(O)R_(f), wherein R_(f) and R_(g) are selected from            hydrogen and (1-4C)alkyl; and        -   dashed lines represent the points of attachment to C and/or            D;

-   W₁, W₂, W₃ and W₄ are independently selected from CR_(h)R_(i),    wherein R_(h) and R_(i) are selected from hydrogen and (1-2C)alkyl;

-   X₁, X₂, X₃ and X₄ are independently selected from a group of the    formula:

-   

-   wherein:    -   

    -   denotes the point of attachment;

    -   W_(x) is selected from O or NH; and

    -   Q is selected from O, S and NR_(j), wherein R_(j) is selected        from hydrogen, (1-4C)alkyl, aryl, heteroaryl and sulfonyl;

-   Z₃ and Z₄ are independently selected from a hydrophilic substituent    group;

-   L is absent or a linker, which optionally bears a hydrophilic    substituent group Z₅;

-   c and d are integers independently selected from 0 to 4; and

-   and p are integers independently selected from 0 t2;

-   and wherein the compound of Formula I is optionally attached to a    displaceable reporter molecule via one or more of the substituent    groups associated with R₁, R₂, R₃, R₄, Z₁, Z₂, Z₃, Z₄ and/or Z₅;

with the proviso that the compound of Formula I comprises at least onehydrophilic substituent group (e.g. Z₁, Z₂, Z₃, Z₄ or Z₅).

Particular compounds of the invention include, for example, compounds ofthe Formula I, or salts, hydrates and/or solvates thereof, wherein,unless otherwise stated, each of bonds b₁ and b₂, Rings A and B, C, D,R₁, R₂, R₃, R₄, W₁, W₂, W₃, W₄, X₁, X₂, X₃, X₄, Z₁, Z₂, Z₃, Z₄, Z₅, L,a, b, c, d, m, n, o, p and any associated substituent groups has any ofthe meanings defined hereinbefore or in any of paragraphs (1) to (60)hereinafter:-

-   (1) bonds b₁ and b₂ are single bonds;

-   (2) bonds b₁ and b₂ are double bonds;

-   (3) R_(1a), R_(1b), R_(2a) and R_(2b) are independently selected    from (1-8C)alkyl, (3-10C)cycloalkyl, aryl, heteroaryl and    heterocyclyl, each of which is optionally substituted by one or more    substituent groups selected from (1-4C)alkyl, halo, (1-4C)haloalkyl,    (1-4C)haloalkoxy, (1-4C)alkoxy, amino, cyano, hydroxyl and a    hydrophilic substituent group; or R_(1a) and R_(1b) are linked so as    to form a group of the formula:

-   

-   -   and/or R_(2a) and R_(2b) are linked so as to form a group of the        formula:

    -   

    -   wherein:        -   

        -   denotes the point of attachment;

        -   bonds b₁ and b₂ are as described above;

        -   Rings A and B are independently selected from aryl,            heteroaryl, heterocyclyl, cycloalkyl and cycloalkenyl;

        -   R₁ and R₂ are independently selected from (1-4C)alkyl, halo,            (1-4C)haloalkyl, (1-4C)haloalkoxy, (1-4C)alkoxy,            (1-4C)alkylamino, amino, cyano, hydroxyl, carboxy,            carbamoyl, sulfamoyl and mercapto;

        -   a and b are integers independently selected from 0 to 2;

        -   m and n are integers independently selected from 0 to 2;

        -   Z₁ and Z₂ are independently selected from hydrophilic            substituent groups;

-   (4) R_(1a), R_(1b), R_(2a) and R_(2b) are independently selected    from aryl and heteroaryl, each of which is optionally substituted by    one or more substituent groups selected from (1-4C)alkyl, halo,    (1-4C)alkoxy, amino or hydroxyl; or    -   R_(1a) and R_(1b) are linked so as to form a group of the        formula:

    -   

    -   and/or R_(2a) and R_(2b) are linked so as to form a group of the        formula:

    -   

    -   wherein:        -   

        -   denotes the point of attachment;

        -   bonds b₁ and b₂ are as described above;

        -   Rings A and B are independently selected from aryl,            heteroaryl, heterocyclyl, cycloalkyl and cycloalkenyl;

        -   R₁ and R₂ are independently selected from (1-4C)alkyl, halo,            (1-4C)haloalkyl, (1-4C)haloalkoxy, (1-4C)alkoxy,            (1-4C)alkylamino, amino, cyano, hydroxyl, carboxy,            carbamoyl, sulfamoyl and mercapto;

        -   a and b are integers independently selected from 0 to 2;

        -   m and n are integers independently selected from 0 to 2;

        -   Z₁ and Z₂ are independently selected from hydrophilic            substituent groups;

-   (5) R_(1a) and R_(1b) are linked so as to form a group of the    formula:

-   

-   -   and R_(2a) and R_(2b) are linked so as to form a group of the        formula:

    -   

    -   wherein:        -   

        -   denotes the point of attachment;

        -   bonds b₁ and b₂ are as described above;

        -   Rings A and B are independently selected from aryl,            heteroaryl, heterocyclyl, cycloalkyl or cycloalkenyl;

        -   R₁ and R₂ are independently selected from (1-4C)alkyl, halo,            (1-4C)haloalkyl, (1-4C)haloalkoxy, (1-4C)alkoxy,            (1-4C)alkylamino, amino, cyano, hydroxyl, carboxy,            carbamoyl, sulfamoyl and mercapto;

        -   a and b are integers independently selected from 0 to 2;

        -   m and n are integers independently selected from 0 to 2;

        -   Z₁ and Z₂ are independently selected from hydrophilic            substituent groups;

-   (6) Rings A and B are independently selected from aryl, heteroaryl    and heterocyclyl (e.g. pyrrolidinyl);

-   (7) Rings A and B are independently selected from aryl and    heteroaryl;

-   (8) Rings A and B are aryl;

-   (9) Rings A and B are independently selected from phenyl, pyridyl,    naphthyl, and pyrrolidinyl;

-   (10) Rings A and B are phenyl or pyrrolidinyl, preferably phenyl;

-   (11) R₁ and R₂ are independently selected from (1-4C)alkyl, halo,    (1-4C)haloalkyl, (1-4C)haloalkoxy, (1-4C)alkoxy, (1-4C)alkylamino,    amino, cyano and hydroxyl;

-   (12) R₁ and R₂ are independently selected from (1-4C)alkyl, halo,    amino, cyano and hydroxyl;

-   (13) C and D are independently selected from aryl, heteroaryl,    heterocyclyl, cycloalkyl, cycloalkenyl and a group of the formula:

-   

-   -   wherein

    -   

    -   denotes the point of attachment;

-   (14) C and D are independently selected from aryl, heteroaryl,    heterocyclyl, cycloalkyl, cycloalkenyl;

-   (15) C and D are independently selected from aryl and heteroaryl;

-   (16) C and D are independently selected from phenyl, naphthenyl and    anthracenyl;

-   (17) C and D are phenyl;

-   (18) C and D are anthracenyl;

-   (19) R₃ and R₄ are independently selected from halo, (1-4C)alkyl,    (1-4C)alkoxy, amino, nitro, (1-4C)alkylamino, (1-4C)dialkylamino,    (1-4C)haloalkyl, (1-4C)haloalkoxy, cyano, (2-4C)alkenyl,    (2-4C)alkynyl and a group of the formula:

-   

-   wherein:    -   L¹ is absent or (1-5C)alkylene;

    -   Y¹ is absent or selected from a one of the following groups; O,        S, SO, SO₂, N(R_(a)), C(O), C(O)O, OC(O), C(O)N(R_(a)),        N(R_(a))C(O), N(R_(b))C(O)N(R_(a)), N(R_(a))C(O)O,        OC(O)N(R_(a)), S(O)₂N(R_(a)), and N(R_(a))SO₂, wherein R_(a) and        R_(b) are each independently selected from hydrogen and        (1-4C)alkyl; and

    -   Q¹ is hydrogen, (1-8C)alkyl, (2-6C)alkenyl, (2-6C)alkynyl, aryl,        (3-10C)cycloalkyl, (3-10C)cycloalkenyl, heteroaryl and        heterocyclyl; wherein

    -   Q¹ is optionally further substituted by one or more substituent        groups independently selected from (1-4C)alkyl, halo,        (1-4C)haloalkyl, (1-4C)haloalkoxy, amino, (1-4C)aminoalkyl,        cyano, hydroxy, carboxy, carbamoyl, sulfamoyl, mercapto, ureido,        oxy, NR_(c)R_(d), OR_(c), C(O)R_(d), C(O)OR_(c), OC(O)R_(c),        C(O)N(R_(d))R_(c), N(R_(d))C(O)R_(c), S(O)_(y)R_(c) (where y is        0, 1 or 2), SO₂N(R_(d))R_(c), N(R_(d))SO₂R_(c),        Si(R_(e))(R_(d))R_(c) and (CH₂)_(z)NR_(d)R_(c) (where z is 1, 2        or 3); wherein R_(c), R_(d) and R_(e) are each independently        selected from hydrogen, (1-6C)alkyl or (3-6C)cycloalkyl; and

    -   two R₃ and/or two R₄ groups taken together may form a group of        the formula:

    -   

    -   wherein:        -   R_(x) is selected from hydrogen and (1-6C)alkyl optionally            substituted by one or more substituent groups selected from            halo, (1-4C)haloalkyl, (1-4C)haloalkoxy, cyano, hydroxy,            sulfamoyl, mercapto, ureido, NR_(f)R_(g), OR_(f), C(O)R_(f),            C(O)OR_(f), OC(O)R_(f), C(O)N(R_(g))R_(f) and            N(R_(g))C(O)R_(f), wherein R_(f) and R_(g) are selected from            hydrogen and (1-4C)alkyl; and        -   dashed lines represent the points of attachment to C and/or            D;

-   (20) R₃ and R₄ are independently selected from halo, (1-4C)alkyl,    (1-4C)alkoxy, amino, nitro, (1-4C)alkylamino, (1-4C)dialkylamino,    (1-4C)haloalkyl, (1-4C)haloalkoxy, cyano, (2-4C)alkenyl,    (2-4C)alkynyl and a group of the formula:

-   

-   wherein:    -   L¹ is absent or a (1-5C)alkylene;

    -   Y¹ is absent or selected from a one of the following groups; O,        S, SO, SO₂, N(R_(a)), C(O), C(O)O, OC(O), C(O)N(R_(a)) and        N(R_(a))C(O), wherein R_(a) is selected from hydrogen and        (1-4C)alkyl; and

    -   Q¹ is hydrogen, (1-8C)alkyl, (2-6C)alkenyl, (2-6C)alkynyl, aryl,        (3-10C)cycloalkyl, (3-10C)cycloalkenyl, heteroaryl and        heterocyclyl; wherein

    -   Q¹ is optionally further substituted by one or more substituent        groups independently selected from (1-4C)alkyl, halo,        (1-4C)haloalkyl, (1-4C)haloalkoxy, amino, (1-4C)aminoalkyl,        cyano, hydroxy, NR_(c)R_(d), OR_(c), C(O)R_(d), C(O)OR_(c),        OC(O)R_(c), C(O)N(R_(d))R_(c) and N(R_(d))C(O)R_(c); wherein        R_(c) and R_(d) are each independently selected from hydrogen        and (1-6C)alkyl; and

    -   two R₃ and/or two R₄ groups taken together may form a group of        the formula:

    -   

    -   wherein:        -   R_(x) is selected from hydrogen and (1-6C)alkyl optionally            substituted by one or more substituent groups selected from            halo, (1-4C)haloalkyl, (1-4C)haloalkoxy, cyano, hydroxy,            sulfamoyl, mercapto, ureido, NR_(f)R_(g), OR_(f), C(O)R_(f),            C(O)OR_(f), OC(O)R_(f), C(O)N(R_(g))R_(f) and            N(R_(g))C(O)R_(f), wherein R_(f) and R_(g) are selected from            hydrogen and (1-4C)alkyl; and        -   dashed lines represent the points of attachment to C and/or            D;

-   (21) R₃ and R₄ are independently selected from halo, (1-4C)alkyl,    (1-4C)alkoxy, amino, nitro, (1-4C)alkylamino, (1-4C)dialkylamino,    (1-4C)haloalkyl, (1-4C)haloalkoxy, cyano, (2-4C)alkenyl,    (2-4C)alkynyl and a group of the formula:

-   

-   wherein:    -   L¹ is absent or (1-5C)alkylene;

    -   Y¹ is absent or selected from a one of the following groups; O,        N(R_(a)), C(O), C(O)O, OC(O), C(O)N(R_(a)) and N(R_(a))C(O),        wherein R_(a) is selected from hydrogen and (1-4C)alkyl; and

    -   Q¹ is hydrogen, (1-8C)alkyl, aryl, (3-10C)cycloalkyl,        (3-10C)cycloalkenyl, heteroaryl and heterocyclyl; wherein Q¹ is        optionally further substituted by one or more substituent groups        independently selected from (1-4C)alkyl, halo, (1-4C)haloalkyl,        (1-4C)haloalkoxy, amino, (1-4C)aminoalkyl, cyano, hydroxy,        NR_(c)R_(d), OR_(c), C(O)R_(d), C(O)OR_(c), OC(O)R_(c),        C(O)N(R_(d))R_(c) and N(R_(d))C(O)R_(c); wherein R_(c) and R_(d)        are each independently selected from hydrogen and (1-6C)alkyl;        and

    -   two R₃ and/or two R₄ groups taken together may form a group of        the formula:

    -   

    -   wherein:        -   R_(x) is selected from hydrogen and (1-6C)alkyl optionally            substituted by one or more substituent groups selected from            halo, (1-4C)haloalkyl, NR_(f)R_(g), OR_(f), C(O)R_(f),            C(O)OR_(f) and C(O)N(R_(g))R_(f), wherein R_(f) and R_(g)            are selected from hydrogen and (1-4C)alkyl; and        -   dashed lines represent the points of attachment to C and/or            D;

-   (22) R₃ and R₄ are independently selected from halo, (1-4C)alkyl,    (1-4C)alkoxy, amino, nitro, (1-4C)alkylamino, (1-4C)dialkylamino,    (1-4C)haloalkyl, (1-4C)haloalkoxy, cyano, (2-4C)alkenyl,    (2-4C)alkynyl and a group of the formula:

-   

-   wherein:    -   L¹ is absent or a (1-5C)alkylene optionally substituted by one        or more substituents selected from (1-2C)alkyl and oxo;    -   Y¹ is absent or selected from a one of the following groups; O,        S, SO, SO₂, N(R_(a)), C(O), C(O)O, OC(O), C(O)N(R_(a)),        N(R_(a))C(O), N(R_(b))C(O)N(R_(a)), N(R_(a))C(O)O,        OC(O)N(R_(a)), S(O)₂N(R_(a)), and N(R_(a))SO₂, wherein R_(a) and        R_(b) are each independently selected from hydrogen and        (1-4C)alkyl; and    -   Q¹ is hydrogen, (1-8C)alkyl, (2-6C)alkenyl, (2-6C)alkynyl, aryl,        (3-10C)cycloalkyl, (3-10C)cycloalkenyl, heteroaryl and        heterocyclyl; wherein    -   Q¹ is optionally further substituted by one or more substituent        groups independently selected from (1-4C)alkyl, halo,        (1-4C)haloalkyl, (1-4C)haloalkoxy, amino, (1-4C)aminoalkyl,        cyano, hydroxy, carboxy, carbamoyl, sulfamoyl, mercapto, ureido,        oxy, NR_(c)R_(d), OR_(c), C(O)R_(d), C(O)OR_(c), OC(O)R_(c),        C(O)N(R_(d))R_(c), N(R_(d))C(O)R_(c), S(O)_(y)R_(c) (where y is        0, 1 or 2), SO₂N(R_(d))R_(c), N(R_(d))SO₂R_(c),        Si(R_(e))(R_(d))R_(c) and (CH₂)_(z)NR_(d)R_(c) (where z is 1, 2        or 3); wherein R_(c), R_(d) and R_(e) are each independently        selected from hydrogen, (1-6C)alkyl and (3-6C)cycloalkyl; and        R_(c) and R_(d) can be linked such that, together with the        nitrogen atom to which they are attached, they form a 4-7        membered heterocyclic ring which is optionally substituted by        one or more substituents selected from (1-4C)alkyl, halo,        (1-4C)haloalkyl, (1-4C)haloalkoxy, (1-4C)alkoxy,        (1-4C)alkylamino, amino, cyano and hydroxyl;

-   (23) R₃ and R₄ are independently selected from halo, (1-4C)alkyl,    (1-4C)alkoxy, amino, nitro, (1-4C)alkylamino, (1-4C)dialkylamino,    (1-4C)haloalkyl, (1-4C)haloalkoxy, cyano, (2-4C)alkenyl,    (2-4C)alkynyl and a group of the formula:

-   

-   wherein:    -   L¹ is absent or (1-5C)alkylene;    -   Y¹ is absent or selected from a one of the following groups; O,        S, SO, SO₂, N(R_(a)), C(O), C(O)O, OC(O), C(O)N(R_(a)) and        N(R_(a))C(O), wherein R_(a) is selected from hydrogen and        (1-4C)alkyl; and    -   Q¹ is hydrogen, (1-8C)alkyl, (2-6C)alkenyl, (2-6C)alkynyl, aryl,        (3-10C)cycloalkyl, (3-10C)cycloalkenyl, heteroaryl and        heterocyclyl; wherein    -   Q¹ is optionally further substituted by one or more substituent        groups independently selected from (1-4C)alkyl, halo,        (1-4C)haloalkyl, (1-4C)haloalkoxy, amino, (1-4C)aminoalkyl,        cyano, hydroxy, carboxy, carbamoyl, sulfamoyl, mercapto, ureido,        oxy, NR_(c)R_(d), OR_(c), C(O)R_(d), C(O)OR_(c), OC(O)R_(c),        C(O)N(R_(d))R_(c), N(R_(d))C(O)R_(c), S(O)_(y)R_(c) (where y is        0, 1 or 2), SO₂N(R_(d))R_(c), N(R_(d))SO₂R_(c),        Si(R_(e))(R_(d))R_(c) and (CH₂)_(z)NR_(d)R_(c) (where z is 1, 2        or 3); wherein R_(c), R_(d) and R_(e) are each independently        selected from hydrogen, (1-6C)alkyl and (3-6C)cycloalkyl;

-   (24) R₃ and R₄ are independently selected from halo, (1-4C)alkyl,    (1-4C)alkoxy, amino, nitro, (1-4C)haloalkyl, (1-4C)haloalkoxy, cyano    and a group of the formula:

-   

-   wherein:    -   L¹ is absent or (1-2C)alkylene;    -   Y¹ is absent or selected from a one of the following groups; O,        N(R_(a)), C(O), C(O)O, OC(O), C(O)N(R_(a)) and N(R_(a))C(O),        wherein R_(a) is selected from hydrogen and (1-4C)alkyl; and    -   Q¹ is hydrogen, (1-8C)alkyl, aryl, (3-10C)cycloalkyl, heteroaryl        and heterocyclyl; wherein Q¹ is optionally further substituted        by one or more substituent groups independently selected from        (1-4C)alkyl, halo, (1-4C)haloalkyl, (1-4C)haloalkoxy, amino,        (1-4C)aminoalkyl, cyano, hydroxy, NR_(c)R_(d), OR_(c),        C(O)R_(d), C(O)OR_(c), OC(O)R_(c), C(O)N(R_(d))R_(c), and        N(R_(d))C(O)R_(c); wherein R_(c) and R_(d) are each        independently selected from hydrogen and (1-6C)alkyl;

-   (25) R₃ and R₄ are independently selected from halo, (1-4C)alkyl,    (1-4C)alkoxy, amino, nitro, (1-4C)haloalkyl, (1-4C)haloalkoxy, cyano    and a group of the formula:

-   

-   wherein:    -   L¹ is absent or (1-2C)alkylene;    -   Y¹ is absent or selected from a one of the following groups; O,        N(R_(a)), C(O)O and C(O)N(R_(a)), wherein R_(a) is selected from        hydrogen and (1-4C)alkyl; and    -   Q¹ is hydrogen, (1-8C)alkyl, aryl and heteroaryl; wherein Q¹ is        optionally further substituted by one or more substituent groups        independently selected from (1-4C)alkyl, halo, (1-4C)haloalkyl,        (1-4C)haloalkoxy, amino, (1-4C)aminoalkyl, cyano, hydroxy,        NR_(c)R_(d), OR_(c), C(O)R_(d), C(O)OR_(c), OC(O)R_(c),        C(O)N(R_(d))R_(c) and N(R_(d))C(O)R_(c); wherein R_(c) and R_(d)        are each independently selected from hydrogen and (1-6C)alkyl;

-   (26) R₃ and R₄ are independently selected from halo, (1-4C)alkyl,    (1-4C)alkoxy, amino, nitro, (1-4C)haloalkyl, (1-4C)haloalkoxy, cyano    and a group of the formula:

-   

-   wherein:    -   L¹ is absent or (1-2C)alkylene;    -   Y¹ is absent or selected from a one of the following groups; O,        N(R_(a)), C(O)O and C(O)N(R_(a)), wherein R_(a) is selected from        hydrogen and (1-4C)alkyl; and    -   Q¹ is hydrogen or (1-8C)alkyl; wherein said (1-8C)alkyl is        optionally further substituted by one or more substituent groups        independently selected from halo, amino, (1-4C)aminoalkyl,        hydroxy, NR_(c)R_(d), OR_(c), C(O)R_(d), C(O)OR_(c) and        C(O)N(R_(d))R_(c); wherein R_(c) and R_(d) are each        independently selected from hydrogen and (1-2C)alky;

-   (27) R₃ and R₄ are independently selected from halo, (1-4C)alkyl,    (1-4C)alkoxy, amino, nitro, (1-4C)haloalkyl, (1-4C)haloalkoxy and    cyano;

-   (28) R₃ and R₄ are independently selected from (1-4C)alkyl and    (1-4C)alkoxy;

-   (29) W₁, W₂, W₃ and W₄ are independently selected from CR_(h)R_(i),    wherein R_(h) and R_(i) are selected from hydrogen and methyl;

-   (30) W₁, W₂, W₃ and W₄ are each CH₂;

-   (31) X₁, X₂, X₃ and X₄ are independently selected from a group of    the formula:

-   

-   wherein:    -   

    -   denotes the point of attachment; and

    -   Q is selected from O, S and NR_(j), wherein R_(j) is selected        from hydrogen, (1-4C)alkyl and aryl;

-   (32) X₁, X₂, X₃ and X₄ are independently selected from a group of    the formula:

-   

-   wherein:    -   

    -   denotes the point of attachment; and

    -   Q is selected from O and S;

-   (33) X₁, X₂, X₃ and X₄ are each a group of the formula:

-   

-   wherein:    -   

    -   denotes the point of attachment;

-   (34) Z₁, Z₂, Z₃, Z₄ and Z₅ are independently selected from a    hydrophilic substituent group, wherein said hydrophilic substituent    comprises one or more hydrophilic functional groups selected from    carboxylic acids, carboxylate ions, carboxylate esters, hydroxyl,    amines, amides, ethers, ketone and aldehyde groups, ureas, nitro    groups, sulphates, sulphonates, phosphates, phosphonates, and    combinations thereof;

-   (35) Z₁, Z₂, Z₃, Z₄ and Z₅ are independently selected from a    hydrophilic substituent group, wherein said hydrophilic substituent    comprises one or more hydrophilic functional groups selected from    carboxylic acids, carboxylate ions, carboxylate esters, hydroxyl,    amines, amides, ethers, ketone groups, aldehyde groups and    combinations thereof;

-   (36) Z₁, Z₂, Z₃, Z₄ and Z₅ are independently selected from a    hydrophilic substituent group, wherein said hydrophilic substituent    comprises one or more hydrophilic functional groups selected from    carboxylic acids, carboxylate ions, hydroxyls, amines and    combinations thereof;

-   (37) Z₁, Z₂, Z₃, Z₄ and Z₅ are independently selected from a    hydrophilic polymer (e.g. polyethylene glycol), a hydrophilic    dendritic group or C(O)OM₁, wherein M₁ is hydrogen or a cation (e.g.    Na, Li, NH₄);

-   (38) Z₁, Z₂, Z₃, Z₄ and Z₅ are independently selected from a    hydrophilic polymer (e.g. polyethylene glycol) or a hydrophilic    dendritic group;

-   (39) Z₁, Z₂, Z₃, Z₄ and Z₅ are independently selected from a    hydrophilic polymer (e.g. polyethylene glycol) or dendritic group    comprising between 1 and 5 generations of building units and a    terminal functional group T₁, wherein each building unit is    independently selected from a group of Formula A:

-   

-   wherein:    -   L² is selected from O, C(O), C(O)O, OC(O), C(O)N(R_(r)),        N(R_(r))C(O), N(R_(s))C(O)N(R_(r)), N(R_(r))C(O)O,        OC(O)N(R_(r)), S(O)₂N(R_(r)), and N(R_(r))SO₂, wherein R_(r) and        R_(s) are each independently selected from hydrogen and        (1-4C)alkyl;

    -   L^(2a) is a bond or a (1-4C)alkylene;

    -   V is absent or a group of the formula:

    -   

    -   

    -   wherein:        -   V₁, V₂, V₃, V₄ and V₅ are independently selected from a            (1-6C)alkylene optionally interrupted by one or more groups            selected from O, S and NR_(t), wherein R_(t) is selected            from hydrogen and (1-2C)alkyl;

        -   #denotes the point of attachment to one of Rings A, B, C, D            or E;

        -   

        -   denotes the point of attachment to either another group of            Formula A or a terminal functional group T₁; and

    -   the terminal functional group T₁ is selected from NH₂, OH,        C(O)OM_(x), C(O)OR_(u) and C(O)NHR_(u), wherein R_(u) is        selected from hydrogen, (1-4C)alkyl, (1-4C)alkoxy,        hydroxy(1-4C)alkyl, (2-4C)alkenyl, (2-4C)alkynyl, ethylene        gylycol and polyethylene glycol, and wherein M_(x) is a cation        (e.g. Na, Li, NH₄);

-   (40) Z₁, Z₂, Z₃, Z₄ and Z₅ are independently selected from a    hydrophilic polymer (e.g. polyethylene glycol) or a dendritic group    comprising between 1 and 5 generations of building units and a    terminal functional group T₁, wherein each building unit is    independently selected from a group of Formula A:

-   

-   wherein:    -   L² is selected from O, C(O), C(O)O, OC(O), C(O)N(R_(r)) and        N(R_(r))C(O), wherein R_(r) is selected from hydrogen and        (1-4C)alkyl;

    -   L^(2a) is a bond or a (1-4C)alkylene;

    -   V is absent or a group of the formula:

    -   

    -   

    -   wherein:        -   V₁, V₂, V₃, V₄ and V₅ are independently selected from a            (1-6C)alkylene optionally interrupted by one or more groups            selected from O, S and NR_(t), wherein R_(t) is selected            from hydrogen and (1-2C)alkyl;

        -   #denotes the point of attachment to one of Rings A, B, C, D            or E;

        -   

        -   denotes the point of attachment to either another group of            Formula A or a terminal functional group T₁; and

    -   the terminal functional group T₁ is selected from OH,        C(O)OM_(x), C(O)OR_(u) and C(O)NHR_(u), wherein R_(u) is        selected from hydrogen, (1-4C)alkyl, (1-4C)alkoxy,        hydroxy(1-4C)alkyl, (2-4C)alkenyl, (2-4C)alkynyl, ethylene        gylycol and polyethylene glycol, and wherein M_(x) is a cation        (e.g. Na, Li, NH₄);

-   (41) Z₁, Z₂, Z₃, Z₄ and Z₅ are independently selected from a    dendritic group comprising between 1 and 4 generations of building    units and a terminal functional group T₁, wherein each building unit    is independently selected from a group of Formula A:

-   

-   wherein:    -   L² is selected from O, C(O), C(O)O, OC(O), C(O)N(R_(r)) and        N(R_(r))C(O), wherein R_(r) is selected from hydrogen and        (1-4C)alkyl;

    -   L^(2a) is a bond or a (1-4C)alkylene;

    -   V is absent or a group of the formula:

    -   

    -   

    -   wherein:        -   V₁, V₂, V₃, V₄ and V₅ are independently selected from a            (1-6C)alkylene optionally interrupted by one or more groups            selected from O and NR_(t), wherein R_(t) is selected from            hydrogen and (1-2C)alkyl;

        -   #denotes the point of attachment to one of Rings A, B, C, D            or E;

        -   

        -   denotes the point of attachment to either another group of            Formula A or a terminal functional group T₁; and

    -   the terminal functional group T₁ is selected from OH,        C(O)OM_(x), C(O)OR_(u) and C(O)NHR_(u), wherein R_(u) is        selected from hydrogen, (1-4C)alkyl, (1-4C)alkoxy and        hydroxy(1-4C)alkyl, wherein M_(x) is a cation (e.g. Na, Li,        NH₄);

-   (42) Z₁, Z₂, Z₃, Z₄ and Z₅ are independently selected from a    hydrophilic polymer or a dendritic group comprising between 1 and 4    generations of building units and a terminal functional group T₁,    wherein each building unit is independently selected from a group of    Formula A:

-   

-   wherein:    -   L² is selected from O, C(O), C(O)O and C(O)N(R_(r)), wherein        R_(r) is selected from hydrogen and (1-4C)alkyl;

    -   L^(2a) is a bond or a (1-4C)alkylene;

    -   V is absent or a group of the formula:

    -   

    -   wherein:        -   V₁, V₂, and V₃ are independently selected from a            (1-6C)alkylene optionally interrupted by one or more groups            selected from oxygen atoms;

        -   #denotes the point of attachment to one of Rings A, B, C, D            or E;

        -   

        -   denotes the point of attachment to either another group of            Formula A or a terminal functional group T₁; and

    -   the terminal functional group T₁ is selected from OH,        C(O)OM_(x), C(O)OR_(u) and C(O)NHR_(u), wherein R_(u) is        selected from hydrogen, (1-4C)alkoxy and hydroxy(1-4C)alkyl,        wherein M_(x) is a cation (e.g. Na, Li, NH₄);

-   (43) Z₁, Z₂, Z₃, Z₄ and Z₅ are independently selected from a    dendritic group comprising between 1 and 4 generations of building    units and a terminal functional group T₁, wherein each building unit    is independently selected from a group of Formula A:

-   

-   wherein:    -   L² is selected from O, C(O)O and C(O)N(R_(r)), wherein R_(r) is        selected from hydrogen and (1-4C)alkyl;

    -   L^(2a) is a bond or a (1-4C)alkylene;

    -   V is a group of the formula:

    -   

    -   wherein:        -   V₁, V₂, and V₃ are independently selected from a            (1-6C)alkylene optionally interrupted by one or more groups            selected from oxygen atoms;

        -   #denotes the point of attachment to one of Rings A, B, C, D            or E;

        -   

        -   denotes the point of attachment to either another group of            Formula A or a terminal functional group T₁; and

    -   the terminal functional group T₁ is selected from OH and        C(O)OM_(x), wherein M_(x) is a cation (e.g. Na, Li, NH₄);

-   (44) Z₁, Z₂, Z₃, Z₄ and Z₅ are independently selected from a    dendritic group comprising between 1 and 3 generations of building    units and a terminal functional group T₁, wherein each building unit    is independently selected from a group of Formula A:

-   

-   wherein:    -   L² is C(O)N(R_(r)), wherein R_(r) is selected from hydrogen and        (1-4C)alkyl;

    -   L^(2a) is a bond or a (1-2C)alkylene;

    -   V is a group of the formula:

    -   

    -   wherein:        -   V₁, V₂, and V₃ are independently selected from a            (1-4C)alkylene optionally interrupted by one or more groups            selected from oxygen atoms;

        -   #denotes the point of attachment to one of Rings A, B, C, D            or E;

        -   

        -   denotes the point of attachment to either another group of            Formula A or a terminal functional group T₁; and

    -   the terminal functional group T₁ is selected is C(O)OM_(x),        wherein M_(x) is a cation (e.g. Na, Li, NH₄);

-   (45) L is absent or a linker of between 8 and 12 atoms in length    (e.g. 10 atoms in length), which optionally bears a hydrophilic    substituent group Z₅;

-   (46) L is absent;

-   (47) L is a linker of between 8 and 12 atoms in length (e.g. 10    atoms in length), which optionally bears a hydrophilic substituent    group Z₅;

-   (48) L is absent or selected from a group of the formula:

-   

-   wherein:    -   

    -   denotes the point of attachment;

    -   W₅ and W₆ are independently selected from CR_(k)R_(l), wherein        R_(k) and R_(l) are selected from hydrogen and (1-2C)alkyl;

    -   X₅ and X₆ are independently selected from a group of the        formula:

    -   

    -   wherein:        -   

        -   denotes the point of attachment; and

        -   Q₂ is selected from O, S and NR_(m), wherein R_(m), is            selected from hydrogen, (1-4C)alkyl, aryl, heteroaryl and            sulfonyl;

    -   bond b₃ is a single or double bond;

    -   Ring E is selected from aryl, heteroaryl, heterocyclyl,        cycloalkyl and cycloalkenyl;

    -   R₅ is selected from (1-4C)alkyl, halo, (1-4C)haloalkyl,        (1-4C)haloalkoxy, (1-4C)alkoxy, (1-4C)alkylamino, amino, cyano,        hydroxyl, carboxy, carbamoyl, sulfamoyl and mercapto;

    -   Z₅ is a hydrophilic substituent group as defined herein;

    -   q is an integer from 0 to 2; and

    -   e is an integer from 0 to 2;

-   (49) L is absent or selected from a group of the formula:

-   

-   wherein:    -   

    -   denotes the point of attachment;

    -   W₅ and W₆ are CH₂;

    -   X₅ and X₆ are independently selected from a group of the        formula:

    -   

    -   wherein:        -   

        -   denotes the point of attachment; and

        -   Q₂ is selected from O or S;

    -   bond b₃ is a single or double bond;

    -   Ring E is selected from aryl and heteroaryl; and

    -   R₅ is selected from (1-4C)alkyl, halo, (1-4C)haloalkyl,        (1-4C)haloalkoxy, (1-4C)alkoxy, (1-4C)alkylamino, amino, cyano,        hydroxyl, carboxy, carbamoyl, sulfamoyl and mercapto;

    -   Z₅ is a hydrophilic substituent group as defined herein;

    -   q is an integer from 0 to 1; and

    -   e is an integer from 0 to 1;

-   (50) L is absent or selected from a group of the formula:

-   

-   wherein:    -   R₅ is selected from (1-4C)alkyl, halo, (1-4C)haloalkyl,        (1-4C)haloalkoxy, (1-4C)alkoxy, (1-4C)alkylamino, amino, cyano,        hydroxyl, carboxy, carbamoyl, sulfamoyl and mercapto;    -   Z₅ is a hydrophilic substituent group as defined herein;    -   q is an integer from 0 to 1; and    -   e is an integer from 0 to 1;

-   (51) L is absent or selected from a group of the formula:

-   

-   wherein:    -   R₅ is selected from (1-4C)alkyl, halo, (1-4C)haloalkyl,        (1-4C)haloalkoxy, (1-4C)alkoxy, (1-4C)alkylamino, amino, cyano,        hydroxyl, carboxy, carbamoyl, sulfamoyl and mercapto;    -   Z₅ is a hydrophilic substituent group as defined herein; and    -   q is 1;

-   (52) c and d are integers independently selected from 0 to 4 (e.g. 0    to 3);

-   (53) c and d are integers independently selected from 1 to 3;

-   (54) one of c and d is 3 and the other is an integer selected from 0    to 3;

-   (55) one of c and d is 3 and the other is an integer selected from 1    to 3;

-   (56) c and d are both 3;

-   (57) a and b are integers independently selected from 0 to 1;

-   (58) a and b are 0;

-   (59) m, n, o and p are integers independently selected from 0 to 1;

-   (60) m and n are 1 and o and p are 0.

In paragraphs (39) to (44) above, the term “generation” will be readilyunderstood to refer to the number of layers of building units (e.g.groups of Formula A) that make up the dendritic group. The term“generation” is a term of the art commonly used in the field ofdendrimer chemistry and will be readily understood by the skilledperson. For example, a one generation dendritic group will be understoodto have one layer (generation) of building units, e.g. -[[buildingunit]]. A two generation dendritic group has two layers of buildingunits, for example, when the building units have trifunctional branchingpoints, the dendritic group may be: -[[building unit][building unit]₃],a three generation dendritric group has three layers of building units,for example -[[building unit][building unit]₃[building unit]₉]. In thisregard, the person skilled in the art will readily appreciate that whenthe dendritic group comprises 1 generation of building units of FormulaA,

denotes the attachment point to the terminal functional group T₁.Moreover, when the dendritic group comprises 2 generations of buildingunits of Formula A, the person skilled in the art will appreciate thatfor the first generation of building units of Formula A,

denotes the attachment point to a second generation of building units ofFormula A, and for the second generation of building units of Formula A,

denotes the attachment point to the terminal functional group T₁.

For illustration purposes only, below there is provided a schematicrepresentations of both a dentritic group comprising one generation ofbuilding units of Formula A and a dendritic group comprising twogenerations of building units of Formula A. In the schematic, Acorresponds to a building unit of Formula A with a trifunctionalbranching point, as described hereinabove, and T₁ corresponds to theterminal functional group T₁, as described hereinabove.

In an embodiment, at least one of integers m, n, o or p is 1 or more. Inthis regard, it will be understood that at least one of Z₁, Z₂, Z₃, Z₄or Z₅ is present.

In another embodiment, a (displaceable) reporter molecule is attached tothe compound of Formula I via one or more of the substituent groups R₁,R₂, R₃, R₄, Z₁, Z₂, Z₃, Z₄ or Z₅. Suitably, the (displaceable) reportermolecule is attached to the compound of Formula I via one or more of thehydrophilic substituent groups Z₁, Z₂, Z₃, Z₄ or Z₅. It will beunderstood that the (displaceable) reporter molecule may be attached toone or more of the substituent groups R₁, R₂, R₃, R₄, Z₁, Z₂, Z₃, Z₄ orZ₅ directly or via a suitable linker (e.g. a polyethylene glycollinker). Suitably, the (displaceable) reporter molecule is attached toone or more of the substituent groups Z₁, Z₂, Z₃, Z₄ or Z₅.

Suitably, the (displaceable) reporter molecule is an aromatic moleculeand/or dye molecule. More suitably, the (displaceable) reporter moleculeis an aromatic molecule, most suitably, a fluorescent aromatic molecule(e.g. fluoresceinamine or tetramethylrhodamine isothiocyanate).

As defined hereinabove, the compound of Formula I may be optionallyattached to a substituent group of Formula A1 at a position associatedwith one or more of the substituent groups R_(1a), R_(1b), R_(2a),R_(2b), R₁, R₂, R₃, R₄, Z₁, Z₂, Z₃, Z₄ and/or Z₅ (e.g. one or more ofthe substitutent groups R₁ or R₂ or one or more of the substituentgroups R₃ or R₄). It will therefore be appreciated that the substituentgroup of Formula A1 may either take the place of the one or moresubstituent groups R_(1a), R_(1b), R_(2a), R_(2b), R₁, R₂, R₃, R₄, Z₁,Z₂, Z₃, Z₄ and/or Z₅, or the substituent group of Formula A1 may beattached to the one or more substituent groups R_(1a), R_(1b), R_(2a),R_(2b), R₁, R₂, R₃, R₄, Z₁, Z₂, Z₃, Z₄ and/or Z₅. Suitably, whenpresent, the substituent group of Formula A1 takes the place of the oneor more substituent groups R_(1a), R_(1b), R_(2a), R_(2b), R₁, R₂, R₃,R₄, Z₁, Z₂, Z₃, Z₄ and/or Z₅.

In an embodiment, the compound of Formula I may be optionally attachedto a substituent group of Formula A1 at a position associated with oneor more of the substituent groups R_(1a), R_(1b), R_(2a), R_(2b), R₁,R₂, R₃ and/or R₄ (e.g. one or more of the substitutent groups R₁ or R₂and/or one or more of the substituent groups R₃ or R₄), wherein thesubstituent group of Formula A1 is of the formula show below:

wherein

-   X_(2a) is absent or selected from O, S, SO, SO₂, N(R^(x2)), C(O),    C(O)O, OC(O), C(O)N(R^(x2)) and N(R^(x2))C(O), wherein R^(x2) is    selected from hydrogen and (1-4C)alkyl;-   L_(2a) is absent or selected from (1-20C)alkylene, (1-20C)alkylene    oxide, (1-20C)alkenyl and (1-20C)alkynyl, each of which being    optionally substituted by one or more substituents selected from    (1-2C)alkyl, aryl and oxo; and-   Z_(2a) is selected from carboxy, carbamoyl, sulphamoyl, mercapto,    amino, azido, (1-4C)alkenyl, (1-4C)alkynyl, NR^(xc)R^(xd), OR^(xc),    ONR^(xc)R^(xd,) C(O)X_(a), C(O)OR^(xf), N=C=O, NR^(xc)C(O)CH₂X_(b),    C(O)N(R^(xe))NR^(Xc)R^(Xd), S(O)_(y)X_(a) (where y is 0, 1 or 2),    SO₂N(R^(xe))NR^(xc)R^(xd), Si(R^(xg))(R^(xh))R^(xi) and an amino    acid; wherein:    -   X_(a) is hydrogen or a leaving group (e.g. halo or CF₃);    -   X_(b) is a halo (e.g. iodo);    -   R^(xc), R^(xd) and R^(xe) are each independently selected from        hydrogen and (1-6C)alkyl;    -   R^(xf) is selected from hydrogen and (1-6C)alkyl, or R^(xf) is a        substituent group that renders C(O)OR^(xf), when taken as a        whole, to be an activated ester (e.g a hydroxysuccinimide ester,        a hydroxy-3-sulfo-succinimide ester or a pentafluorophenyl        ester);    -   R^(xg), R^(xh) and R^(xi) are each independently selected from        (1-4C)alkyl, hydroxy, halo and (1-4C)alkoxy.

In another embodiment, the compound of Formula I may be optionallyattached to a substituent group of Formula A1 at a position associatedwith one or more of the substituent groups R_(1a), R_(1b), R_(2a),R_(2b), R₁, R₂, R₃ and/or R₄, wherein the substituent group of FormulaA1 is of the formula show below:

wherein

-   X_(2a) is absent or selected from O, S, SO, SO₂, N(R^(x2)), C(O),    C(O)O, OC(O), C(O)N(R^(x2)) and N(R^(x2))C(O), wherein R^(x2) is    selected from hydrogen and (1-4C)alkyl;-   L_(2a) is absent or selected from (1-20C)alkylene, (1-20C)alkylene    oxide, (1-20C)alkenyl and (1-20C)alkynyl, each of which being    optionally substituted by one or more substituents selected from    (1-2C)alkyl, aryl and oxo; and-   Z_(2a) is selected from carboxy, carbamoyl, sulphamoyl, mercapto,    amino, azido, (1-4C)alkenyl, (1-4C)alkynyl, NR^(xc)R^(xd), OR^(xc),    C(O)X_(a), C(O)OR^(xf), N=C=O, NR^(xc)C(O)CH₂X_(b) and    C(O)N(R^(xe))NR^(Xc)R^(Xd); wherein:    -   X_(a) is hydrogen or a leaving group (e.g. halo or CF₃);    -   X_(b) is a halo (e.g. iodo);    -   R^(xc), R^(xd) and R^(xe) are each independently selected from        hydrogen and (1-6C)alkyl;    -   R^(xf) is selected from hydrogen and (1-6C)alkyl, or R^(xf) is a        substituent group that renders C(O)OR^(xf), when taken as a        whole, to be an activated ester (e.g a hydroxysuccinimide ester,        a hydroxy-3-sulfo-succinimide ester or a pentafluorophenyl        ester);    -   R^(xg), R^(xh) and R^(xi) are each independently selected from        (1-4C)alkyl, hydroxy, halo and (1-4C)alkoxy.

In a further embodiment, the compound of Formula I may be optionallyattached to a substituent group of Formula A1 at a position associatedwith one or more of the substituent groups R_(1a), R_(1b), R_(2a),R_(2b), R₁, R₂, R₃ and/or R₄, wherein the substituent group of FormulaA1 is of the formula show below:

wherein

-   X_(2a) is absent or selected from O, N(R^(x2)), C(O)O, OC(O),    C(O)N(R^(x2)) and N(R^(x2))C(O), wherein R^(x2) is selected from    hydrogen and (1-4C)alkyl;-   L_(2a) is absent or selected from (1-20C)alkylene, (1-20C)alkylene    oxide, (1-20C)alkenyl and (1-20C)alkynyl, each of which being    optionally substituted by one or more substituents selected from    (1-2C)alkyl and oxo; and-   Z_(2a) is selected from carboxy, carbamoyl, sulphamoyl, mercapto,    amino, azido, (1-4C)alkenyl, (1-4C)alkynyl, NR^(xc)R^(xd), OR^(xc),    C(O)OR^(xf) and N=C=O; wherein:    -   R^(xc) and R^(xd) are each independently selected from hydrogen        and (1-6C)alkyl; and    -   R^(xf) is selected from hydrogen and (1-6C)alkyl, or R^(xf) is a        substituent group that renders C(O)OR^(xf), when taken as a        whole, to be an activated ester (e.g a hydroxysuccinimide ester,        a hydroxy-3-sulfo-succinimide ester or a pentafluorophenyl        ester).

In yet a further embodiment, the compound of Formula I may be optionallyattached to a substituent group of Formula A1 at a position associatedwith one or more of the substituent groups R_(1a), R_(1b), R_(2a),R_(2b), R₁, R₂, R₃ and/or R₄, wherein the substituent group of FormulaA1 is of the formula show below:

wherein

-   X_(2a) is absent or selected from O, N(R^(x2)), C(O)O and    C(O)N(R^(x2)), wherein R^(x2) is selected from hydrogen and    (1-4C)alkyl;-   L_(2a) is absent or selected from (1-10C)alkylene, (1-10C)alkylene    oxide, (1-10C)alkenyl and (1-10C)alkynyl, each of which being    optionally substituted by one or more substituents selected from    (1-2C)alkyl and oxo; and-   Z_(2a) is selected from carboxy, carbamoyl, sulphamoyl, mercapto,    amino, azido, (1-4C)alkenyl, (1-4C)alkynyl, NR^(xc)R^(xd,) OR^(xc)    C(O)OR^(xf) and N=C=O; wherein:    -   R^(xc) and R^(xd) are each independently selected from hydrogen        and (1-6C)alkyl; and    -   R^(xf) is selected from hydrogen, (1-6C)alkyl, succinimide,        3-sulfo-succinimide and pentafluorophenyl.

Suitably, a heteroaryl or heterocyclyl group as defined herein is amonocyclic heteroaryl or heterocyclyl group comprising one, two or threeheteroatoms selected from N, O or S.

Suitably, a heteroaryl is a 5- or 6-membered heteroaryl ring comprisingone, two or three heteroatoms selected from N, O or S.

Suitably, a heterocyclyl group is a 4-, 5- or 6-membered heterocyclylring comprising one, two or three heteroatoms selected from N, O or S.Most suitably, a heterocyclyl group is a 5-, 6- or 7-membered ringcomprising one, two or three heteroatoms selected from N, O or S [e.g.morpholinyl (e.g. 4-morpholinyl), pyridinyl, piperazinyl,homopiperazinyl or pyrrolidinonyl].

Suitably an aryl group is phenyl or anthracenyl, most suitably phenyl.

Suitably, bonds b₁ and b₂ are as defined in any one of paragraphs (1) or(2) above.

Suitably, R_(1a), R_(1b), R_(2a) and R_(2b) are as defined in any one ofparagraphs (3) to (5) above. Most suitably, R_(1a), R_(1b), R_(2a) andR_(2b) are as defined in paragraph (5) above.

Suitably, Rings A and B are as defined in any one of paragraphs (6) to(10) above. Most suitably, Rings A and B are phenyl.

Suitably, R₁ and R₂ are as defined in any one of paragraphs (11) to (12)above.

Suitably, C and D are as defined in any one of paragraphs (13) to (18)above. Most suitably, C and D are as defined in any one of paragraphs(17) or (18) above.

Suitably, R₃ and R₄ are as defined in any one of paragraphs (19) to (28)above. Most suitably, R₃ and R₄ are as defined in paragraph (28) above.

Suitably, W₁, W₂, W₃ and W₄ are as defined in any one of paragraphs (29)or (30) above.

Suitably, X₁, X₂, X₃ and X₄ are as defined in any one of paragraphs (31)to (33) above. Most suitably, X₁, X₂, X₃ and X₄ are as defined inparagraph (33) above.

Suitably, Z₁, Z₂, Z₃, Z₄ and Z₅ are as defined in any one of paragraphs(34) to (44) above. Most suitably, Z₁, Z₂, Z₃, Z₄ and Z₅ are as definedin paragraph (44) above.

Suitably, L is as defined in any one of paragraphs (45) to (51) above.

Suitably, integres c and d are as defined in any one of paragraphs (52)to (56).

Suitably, integers a and b are as defined in any one of paragraphs (57)to (58) above.

Suitably, integers m, n, o and p are as defined in any one of paragraphs(59) to (60) above.

In a particular group of compounds of the invention, R_(1a) and R_(1b)together with R_(2a) and R_(2b) are linked to form Rings A and Brespectively, i.e. the compounds have the structural formula la (asub-definition of Formula (I)) shown below, or a salt, hydrate and/orsolvate thereof:

wherein, each of bonds b₁ and b₂, R₁, R₂, R₃, R₄, Z₁, Z₂, Z₃, Z₄, a, b,c, d, m, n, o, p, L, C, D and Rings A and B, are as defined hereinabove.

In an embodiment of the compounds of Formula la:

-   bonds b₁ and b₂ are as defined in any one of paragraphs (1) or (2)    above;-   Rings A and B are as defined in any one of paragraphs (6) to (10)    above;-   R₁ and R₂ are as defined in any one of paragraphs (11) to (12)    above;-   C and D are as defined in any one of paragraphs (13) to (18) above;-   R₃ and R₄ are as defined in any one of paragraphs (19) to (28)    above;-   W₁, W₂, W₃ and W₄ are as defined in any one of paragraphs (29)    to (30) above;-   X₁, X₂, X₃ and X₄ are as defined in any one of paragraphs (31)    to (33) above;-   Z₁, Z₂, Z₃, Z₄ and Z₅ are as defined in any one of paragraphs (34)    to (44) above;-   L is as defined in any one of paragraphs (45) to (51) above;-   intergers c and d are as defined in any one of paragraphs (52)    to (56) above-   intergers a and b are as defined in any one of paragraphs (57)    to (58) above; and-   integers m, n, o and p are as defined in any one of paragraphs (59)    to (60) above.

In another embodiment of the compounds of Formula la:

-   bonds b₁ and b₂ are as defined in paragraph (2) above;-   Rings A and B are as defined in paragraph (10) above;-   R₁ and R₂ are as defined in paragraphs (12) above;-   C and D are as defined in any one of paragraphs (17) or (18) above;-   R₃ and R₄ are as defined in paragraph (28) above;-   W₁, W₂, W₃ and W₄ are as defined in paragraph (30) above;-   X₁, X₂, X₃ and X₄ are as defined in paragraph (33) above;-   Z₁, Z₂, Z₃, Z₄ and Z₅ are as defined in any one of paragraphs (42)    to (44) above;-   L is as defined in paragraphs (51) above;-   intergers c and d are as defined in any one of paragraphs (52)    to (56) above;-   intergers a and b are as defined in any one of paragraphs (57)    to (58) above; and-   integers m, n, o and p are as defined in any one of paragraphs (59)    to (60) above.

In a particular group of compounds of the invention, R_(1a) and R_(1b)together with R_(2a) and R_(2b) are linked to form Rings A and Brespectively, Q is O and W₁, W₂, W₃ and W₄ are CH₂, i.e. the compoundshave the structural formula Ib (a sub-definition of Formula (I)) shownbelow, or a salt, hydrate and/or solvate thereof:

wherein, each of bonds b₁ and b₂, R₁, R₂, R₃, R₄, Z₁, Z₂, Z₃, Z₄, a, b,c, d, m, n, o, p, L, C, D and Rings A and B, are as defined hereinabove.

In an embodiment of the compounds of Formula Ib:

-   bonds b₁ and b₂ are as defined in any one of paragraphs (1) or (2)    above;-   Rings A and B are as defined in any one of paragraphs (6) to (10)    above;-   R₁ and R₂ are as defined in any one of paragraphs (11) to (12)    above;-   C and D are as defined in any one of paragraphs (13) to (18) above;-   R₃ and R₄ are as defined in any one of paragraphs (19) to (28)    above;-   Z₁, Z₂, Z₃, Z₄ and Z₅ are as defined in any one of paragraphs (34)    to (44) above;-   L is as defined in any one of paragraphs (45) to (51) above;-   intergers c and d are as defined in any one of paragraphs (52)    to (56) above;-   intergers a and b are as defined in any one of paragraphs (57)    to (58) above; and-   integers m, n, o and p are as defined in any one of paragraphs (59)    to (60) above.

In another embodiment of the compounds of Formula Ib:

-   bonds b₁ and b₂ are as defined in paragraph (2) above;-   Rings A and B are as defined in paragraph (10) above;-   R₁ and R₂ are as defined in paragraphs (12) above;-   C and D are as defined in any one of paragraphs (17) or (18) above;-   R₃ and R₄ are as defined in paragraph (28) above;-   Z₁, Z₂, Z₃, Z₄ and Z₅ are as defined in any one of paragraphs (40)    to (44) above;-   L is as defined in paragraphs (51) above;-   intergers c and d are as defined in any one of paragraphs (52)    to (56) above;-   intergers a and b are as defined in any one of paragraphs (57)    to (58) above; and-   integers m, n, o and p are as defined in any one of paragraphs (59)    to (60) above.

In another particular group of compounds of the invention, R_(1a) andR_(1b) together with R_(2a) and R_(2b) are linked to form Rings A and Brespectively, Q is O, W₁, W₂, W₃ and W₄ are CH₂, and L is as shownbelow, i.e. the compounds have the structural formula Ic (asub-definition of Formula (I)) shown below, or a salt, hydrate and/orsolvate thereof:

wherein, each of bonds b₁, b₂ and b₃, R₁, R₂, R₃, R₄, R₅, Z₁, Z₂, Z₃,Z₄, Z₅, a, b, c, d, e, m, n, o, p, q, C, D and Rings A, B and E are asdefined hereinabove.

In an embodiment of the compounds of Formula Ic:

-   bonds b₁ and b₂ are as defined in any one of paragraphs (1) or (2)    above;-   Rings A and B are as defined in any one of paragraphs (6) to (10)    above;-   R₁ and R₂ are as defined in any one of paragraphs (11) to (12)    above;-   C and D are as defined in any one of paragraphs (13) to (18) above;-   R₃ and R₄ are as defined in any one of paragraphs (19) to (28)    above;-   Z₁, Z₂, Z₃, Z₄ and Z₅ are as defined in any one of paragraphs (34)    to (44) above;-   bond b₃, Ring E, R₅ and intergers e and q are as defined in any one    of paragraphs (48) to (49) above;-   intergers c and d are as defined in any one of paragraphs (52)    to (56) above;-   intergers a and b are as defined in any one of paragraphs (57)    to (58) above; and-   integers m, n, o and p are as defined in any one of paragraphs (59)    to (60) above.

In another embodiment of the compounds of Formula Ic:

-   bonds b₁ and b₂ are as defined in paragraph (2) above;-   Rings A and B are as defined in paragraph (10) above;-   R₁ and R₂ are as defined in paragraph (12) above;-   C and D are as defined in any one of paragraphs (17) to (18) above;-   R₃ and R₄ are as defined in paragraph (28) above;-   Z₁, Z₂, Z₃, Z₄ and Z₅ are as defined in any one of paragraphs (40)    to (44) above;-   bond b₃, Ring E, R₅ and intergers e and q are as defined in    paragraph (49) above;-   intergers c and d are as defined in any one of paragraphs (52)    to (56) above;-   intergers a and b are as defined in any one of paragraphs (57)    to (58) above; and-   integers m, n, o and p are as defined in any one of paragraphs (59)    to (60) above.

In yet another particular group of compounds of the invention, Q is O,W₁, W₂, W₃ and W₄ are CH₂, and L is as shown below, Rings A, B and E arephenyl, integers m and n are 1 and integers a, b and e are 0, i.e. thecompounds have the structural formula Id (a sub-definition of Formula(I)) shown below, or a salt, hydrate and/or solvate thereof:

wherein, each of R₃, R₄, Z₁, Z₂, Z₃, Z₄, Z₅, c, d, o, p and Rings C andD are as defined hereinabove.

In an embodiment of the compounds of Formula Id:

-   Rings C and D are as defined in any one of paragraphs (13) to (18)    above;-   R₃ and R₄ are as defined in any one of paragraphs (19) to (28)    above;-   Z₁, Z₂, Z₃, Z₄ and Z₅ are as defined in any one of paragraphs (34)    to (44) above;-   intergers c and d are as defined in any one of paragraphs (42)    to (56) above; and-   integers o and p are as defined in any one of paragraphs (59)    to (60) above.

In another particular group of compounds of the invention, Q is O, W₁,W₂, W₃ and W₄ are CH₂, and L is as shown below, Rings A, B and E arephenyl, integers m and n are 1 and integers a, b and e are 0, i.e. thecompounds have the structural formula le (a sub-definition of Formula(I)) shown below, or a salt, hydrate and/or solvate thereof:

wherein, each of Z₁, Z₂, Z₃, Z₄ and Z₅ are as defined hereinabove andR^(3a), R^(3b), R^(3c) _(,) R^(4a), R^(4b) and R^(4c) are independentlyselected from hydrogen, halo, (1-4C)alkyl, (1-4C)alkoxy, amino, nitro,(1-4C)alkylamino, (1-4C)dialkylamino, (1-4C)haloalkyl, (1-4C)haloalkoxy,cyano, (2-4C)alkenyl, (2-4C)alkynyl and a group of the formula:

wherein:

-   L^(1a) is absent or (1-2C)alkylene optionally substituted by one or    more substituents selected from (1-2C)alkyl and oxo;-   Y^(1a) is absent or O, S, SO, SO₂, N(R_(n)), C(O), C(O)O, OC(O),    C(O)N(R_(n)) and N(R_(n))C(O), wherein R_(n) is selected from    hydrogen and (1-4C)alkyl; and-   Q^(1a) is hydrogen, (1-8C)alkyl, (2-6C)alkenyl, (2-6C)alkynyl, aryl,    (3-10C)cycloalkyl, (3-10C)cycloalkenyl, heteroaryl and heterocyclyl;    wherein-   Q^(1a) is optionally further substituted by one or more substituent    groups independently selected from (1-4C)alkyl, halo,    (1-4C)haloalkyl, (1-4C)haloalkoxy, amino, (1-4C)aminoalkyl, cyano,    hydroxy, carboxy, carbamoyl, sulfamoyl, mercapto, ureido, oxy,    NR_(o)R_(p), OR_(o), C(O)R_(o), C(O)OR_(o), OC(O)R_(o),    C(O)N(R_(p))R_(o), N(R_(p))C(O)R_(o), S(O)_(y1) R_(o) (where y₁ is    0, 1 or 2), SO₂N(R_(p))R_(o), N(R_(p))SO₂R_(o),    Si(R_(q))(R_(p))R_(o) and (CH₂)_(z1)NR_(o)R_(p) (where z₁ is 1, 2 or    3); wherein R_(o), R_(p) and R_(q) are each independently selected    from hydrogen and (1-6C)alkyl.

In an embodiment of the compounds of Formula le:

-   Z₁, Z₂, Z₃, Z₄ and Z₅ are as defined in any one of paragraphs (34)    to (42) above; and R^(3a), R^(3b), R^(3c) _(,) R^(4a), R^(4b) and    R^(4c) are independently selected from hydrogen, halo, (1-4C)alkyl,    (1-4C)alkoxy, amino, nitro, (1-4C)alkylamino, (1-4C)dialkylamino,    (1-4C)haloalkyl, (1-4C)haloalkoxy, cyano, (2-4C)alkenyl,    (2-4C)alkynyl and a group of the formula:

-   

-   wherein:    -   L^(1a) is absent or (1-2C)alkylene;    -   Y^(1a) is absent or O, S, SO, SO₂, N(R_(l)), C(O), C(O)O, OC(O),        C(O)N(R_(n)) and N(R_(n))C(O), wherein R_(n) is selected from        hydrogen and (1-4C)alkyl; and    -   Q^(1a) is hydrogen, (1-8C)alkyl, (2-6C)alkenyl, (2-6C)alkynyl,        aryl, (3-10C)cycloalkyl, (3-10C)cycloalkenyl, heteroaryl or        heterocyclyl; wherein    -   Q^(1a) is optionally further substituted by one or more        substituent groups independently selected from (1-4C)alkyl,        halo, (1-4C)haloalkyl, (1-4C)haloalkoxy, amino,        (1-4C)aminoalkyl, cyano, hydroxy, carboxy, carbamoyl, sulfamoyl        and mercapto

In another embodiment of the compounds of Formula le:

-   Z₁, Z₂, Z₃, Z₄ and Z₅ are as defined in paragraph (42) above; and-   R^(3a), R^(3b), R^(3c) _(,) R^(4a), R^(4b) and R^(4c) are    independently selected from hydrogen, halo, (1-4C)alkyl,    (1-4C)alkoxy, amino, nitro, (1-4C)alkylamino, (1-4C)dialkylamino,    (1-4C)haloalkyl, (1-4C)haloalkoxy, cyano, (2-4C)alkenyl and    (2-4C)alkynyl.

In another embodiment of the compounds of Formula le:

-   Z₁, Z₂, Z₃, Z₄ and Z₅ are as defined in paragraph (42) above; and-   R^(3a), R^(3b), _(R) ^(3c) _(,) R^(4a), R^(4b) and R^(4c) are    independently selected from hydrogen, halo, (1-4C)alkyl,    (1-4C)alkoxy, amino, nitro, (1-4C)alkylamino, (1-4C)dialkylamino,    (1-4C)haloalkyl, (1-4C)haloalkoxy, cyano, (2-4C)alkenyl and    (2-4C)alkynyl,-   with the proviso that R^(3a), R^(3b) and R^(3c) cannot all be    hydrogen.

In another embodiment of the compounds of Formula le:

-   Z₁, Z₂, Z₃, Z₄ and Z₅ are as defined in paragraph (42) above; and-   R^(3a), R^(3b), R^(3c) _(,) R^(4a), R^(4b) and R^(4c) are    independently selected from halo, (1-4C)alkyl, (1-4C)alkoxy, amino,    nitro, (1-4C)alkylamino, (1-4C)dialkylamino, (1-4C)haloalkyl,    (1-4C)haloalkoxy, cyano, (2-4C)alkenyl and (2-4C)alkynyl.

In another embodiment of the compounds of Formula le:

-   Z₁, Z₂, Z₃, Z₄ and Z₅ are as defined in paragraph (44) above; and-   R^(3a), R^(3b), R^(3c) _(,) R^(4a), R^(4b) and R^(4c) are    independently selected from hydrogen and (1-4C)alkyl.

In another embodiment of the compounds of Formula le:

-   Z₁, Z₂, Z₃, Z₄ and Z₅ are as defined in paragraph (44) above; and-   R^(3a), R^(3b), R^(3c) _(,) R^(4a), R^(4b) and R^(4c) are    independently selected from hydrogen, (1-4C)alkoxy and (1-4C)alkyl.

In another embodiment of the compounds of Formula le:

-   Z₁, Z₂, Z₃, Z₄ and Z₅ are as defined in paragraph (44) above; and-   R^(3a), R^(3b), R^(3c) _(,) R^(4a), R^(4b) and R^(4c) are    independently selected from (1-4C)alkoxy and (1-4C)alkyl.

In another embodiment of the compounds of Formula le:

-   Z₁, Z₂, Z₃, Z₄ and Z₅ are as defined in paragraph (44) above; and-   R^(3a), R^(3b), R^(3c) _(,) R^(4a), R^(4b) and R^(4c) are    independently a (1-4C)alkyl (e.g. ethyl).

In yet a further group of compounds of the invention, R_(1a) and R_(1b)together with R_(2a) and R_(2b) are linked to form Rings A and Brespectively, Q is O, L is absent and W₁, W₂, W₃ and W₄ are CH₂, i.e.the compounds have the structural formula If (a sub-definition ofFormula (I)) shown below, or a salt, hydrate and/or solvate thereof:

wherein, each of bonds b₁ and b₂, R₁, R₂, R₃, R₄, Z₁, Z₂, Z₃, Z₄, a, b,c, d, m, n, o, p, C, D and Rings A and B are as defined hereinabove.

In an embodiment of the compounds of Formula If:

-   bonds b₁ and b₂ are as defined in any one of paragraphs (1) or (2)    above;-   Rings A and B are as defined in any one of paragraphs (6) to (10)    above;-   R₁ and R₂ are as defined in any one of paragraphs (11) to (12)    above;-   C and D are as defined in any one of paragraphs (13) to (18) above;-   R₃ and R₄ are as defined in any one of paragraphs (19) to (28)    above;-   Z₁, Z₂, Z₃ and Z₄ are as defined in any one of paragraphs (34)    to (44) above;-   intergers c and d are as defined in any one of paragraphs (52)    to (56) above;-   intergers a and b are as defined in any one of paragraphs (57)    to (58) above; and-   integers m, n, o and p are as defined in any one of paragraphs (59)    to (60) above.

In another embodiment of the compounds of Formula If:

-   bonds b₁ and b₂ are as defined in paragraph (2) above;-   Rings A and B are as defined in paragraph (10) above;-   R₁ and R₂ are as defined in paragraph (12) above;-   C and D are as defined in any one of paragraphs (17) to (18) above;-   R₃ and R₄ are as defined in paragraph (28) above;-   Z₁, Z₂, Z₃ and Z₄ are as defined in any one of paragraphs (38)    to (44) above;-   intergers c and d are as defined in any one of paragraphs (52)    to (56) above;-   intergers a and b are as defined in any one of paragraphs (57)    to (58) above; and-   integers m, n, o and p are as defined in any one of paragraphs (52)    to (53) above.

In aother particular group of compounds of the invention, Q is O, L isabsent, W₁, W₂, W₃ and W₄ are CH₂, Rings A and B are phenyl and Rings Cand D are anthracenyl, i.e. the compounds have the structural formula Ig(a sub-definition of Formula (I)) shown below, or a salt, hydrate and/orsolvate thereof:

wherein each of R₁, R₂, R₃, R₄, Z₁, Z₂, Z₃, Z₄, a, b, c, d, m, n, o andp are as defined hereinabove.

In an embodiment of the compounds of Formula Ig:

-   R₁ and R₂ are as defined in any one of paragraphs (11) to (12)    above;-   R₃ and R₄ are as defined in any one of paragraphs (19) to (28)    above;-   Z₁, Z₂, Z₃ and Z₄ are as defined in any one of paragraphs (34)    to (44) above;-   intergers c and d are as defined in paragraphs (52) above;-   intergers a and b are as defined in any one of paragraphs (57)    to (58) above; and-   integers m, n, o and p are as defined in any one of paragraphs (59)    to (60) above.

Particular compounds of the present invention include any of thecompounds exemplified in the present application, or a salt, solvate orhydrate thereof, and, in particular, any of the following:

-   i)

-   

-   ii)

-   

-   iii)

-   

-   iv)

-   

-   v)

-   

-   vi)

-   

-   vii)

-   

-   viii)

-   

-   ix)

-   

-   x)

-   

-   xi)

-   

-   xii)

-   

-   xiii)

-   

-   xiv)

-   

-   XV)

-   

-   xvi)

-   

-   xvii)

-   

-   xviii)

-   

-   xix)

-   

-   wherein:    -   each of Z₁, Z₂ and Z₅ are independently selected from one of the        following groups:

    -   

    -   

    -   

    -   

    -   wherein each R^(z1) is independently selected from hydrogen or        Na (i.e. the carboxy group is either a carboxylic acid or a        sodium salt thereof);

    -   each of Z₃ and Z₄ is a group of the formula:

    -   

    -   Z¹⁰⁰ is a group of the formula:

    -   

    -   R₁ and R₂ are groups of the formula:

    -   

    -   R^(3a) and R^(4a) are independently selected from hydrogen or        methoxy;

    -   and wherein

    -   

    -   denotes the point of attachment.

Particular compounds of the present invention include any of thecompounds exemplified in the present application, or a salt, solvate orhydrate thereof, and, in particular, any of the following:

-   i)

-   

-   ii)

-   

-   iii)

-   

-   iv)

-   

-   v)

-   

-   vi)

-   

-   vii)

-   

-   viii)

-   

-   ix)

-   

-   x)

-   

-   xi)

-   

-   xii)

-   

-   xiii)

-   

-   wherein:    -   each of Z₁, Z₂ and Z₅ are independently selected one of the        following groups:

    -   

    -   

    -   each of Z₃ and Z₄ is a group of the formula:

    -   

    -   Z¹⁰⁰ is a group of the formula:

    -   

    -   R^(3a) and R^(4a) are independently selected from hydrogen or        methoxy;

    -   and wherein

    -   

    -   denotes the point of attachment.

Further particular compounds of the present invention include any of thecompounds exemplified in the present application, or a salt, solvate orhydrate thereof, and, in particular, any of the following:

Further particular compounds of the present invention include any of thecompounds exemplified in the present application, or a salt, solvate orhydrate thereof, and, in particular, any of the following:

-   i)

-   

-   -   wherein each of Z₁, Z₂ and Z₅ is a group of the formula:

    -   

    -   wherein

    -   

    -   denotes the point of attachment;

-   ii)

-   

-   -   wherein each of Z₁, and Z₂ is a group of the formula:

    -   

    -   wherein

    -   

    -   denotes the point of attachment;

-   iii)

-   

-   -   wherein each of Z₃, and Z₄ is a group of the formula:

    -   

    -   wherein

    -   

    -   denotes the point of attachment;

-   iv)

-   

-   -   wherein each of Z₁, Z₂ and Z₅ is a group of the formula:

    -   

    -   wherein

    -   

    -   denotes the point of attachment; or

-   v)

-   

-   -   wherein each of Z₂ and Z₅ is a group of the formula:

    -   

    -   Z¹⁰⁰ is a group of the formula:

    -   

    -   and wherein

    -   

    -   denotes the point of attachment.

Yet further particular compounds of the present invention include any ofthe compounds exemplified in the present application, or a salt, solvateor hydrate thereof, and, in particular, any of the following:

-   i)

-   

-   -   wherein each of Z₁, Z₂ and Z₅ is a group of the formula:

    -   

    -   wherein

    -   

    -   denotes the point of attachment;

-   ii)

-   

-   -   wherein each of Z₁, and Z₂ is a group of the formula:

    -   

    -   wherein

    -   

    -   denotes the point of attachment;

-   iii)

-   

-   -   wherein each of Z₃, and Z₄ is a group of the formula:

    -   

    -   wherein

    -   

    -   denotes the point of attachment.

Suitably, the compound of the present invention is as follows:

-   wherein each of Z₁, Z₂ and Z₅ is a group of the formula:

-   

-   wherein

-   

-   denotes the point of attachment.

In certain embodiments of the present invention, the compound is not thefollowing compound:

The solid and dashed lines used in Formulae I, la, Ib, Ic, Id, le, Ifand Ig hereinabove will be readily understood to have been used forillustration purposes only (i.e. to display the relative orientation ofthe compounds of the present invention). They do not refer to theabsolute configuration (i.e. stereochemistry) of the compounds shown.

A suitable salt of a compound of the invention is, for example, anacid-addition salt of a compound of the invention which is sufficientlybasic, for example, an acid-addition salt with, for example, aninorganic or organic acid, for example hydrochloric, hydrobromic,sulfuric, phosphoric, trifluoroacetic, formic, citric methane sulfonateor maleic acid. In addition, a suitable salt of a compound of theinvention which is sufficiently acidic is an alkali metal salt, forexample a sodium or potassium salt, an alkaline earth metal salt, forexample a calcium or magnesium salt, an ammonium salt or a salt with anorganic base which affords an acceptable cation, for example a salt withmethylamine, dimethylamine, trimethylamine, piperidine, morpholine ortris-(2-hydroxyethyl)amine.

Compounds that have the same molecular formula but differ in the natureor sequence of bonding of their atoms or the arrangement of their atomsin space are termed “isomers”. Isomers that differ in the arrangement oftheir atoms in space are termed “stereoisomers”. Stereoisomers that arenot mirror images of one another are termed “diastereomers” and thosethat are non-superimposable mirror images of each other are termed“enantiomers”. When a compound has an asymmetric center, for example, itis bonded to four different groups, a pair of enantiomers is possible.An enantiomer can be characterized by the absolute configuration of itsasymmetric center and is described by the R- and S-sequencing rules ofCahn and Prelog, or by the manner in which the molecule rotates theplane of polarized light and designated as dextrorotatory orlevorotatory (i.e., as (+) or (-)-isomers respectively). A chiralcompound can exist as either individual enantiomer or as a mixturethereof. A mixture containing equal proportions of the enantiomers iscalled a “racemic mixture”.

The compounds of this invention may possess one or more asymmetriccenters; such compounds can therefore be produced as individual (R)- or(S)-stereoisomers or as mixtures thereof. Unless indicated otherwise,the description or naming of a particular compound in the specificationand claims is intended to include both individual enantiomers andmixtures, racemic or otherwise, thereof. The methods for thedetermination of stereochemistry and the separation of stereoisomers arewell-known in the art (see discussion in Chapter 4 of “Advanced OrganicChemistry”, 4th edition J. March, John Wiley and Sons, New York, 2001),for example by synthesis from optically active starting materials or byresolution of a racemic form. Some of the compounds of the invention mayhave geometric isomeric centres (E- and Z- isomers). It is to beunderstood that the present invention encompasses all optical,diastereoisomers and geometric isomers and mixtures thereof that arecapable of saccharide recognition.

The present invention also encompasses compounds of the invention asdefined herein which comprise one or more isotopic substitutions. Forexample, H may be in any isotopic form, including 1H, 2H(D), and 3H (T);C may be in any isotopic form, including 12C, 13C, and 14C; and O may bein any isotopic form, including 16O and18O; and the like.

It is also to be understood that certain compounds of the Formula (I),and sub-formulae la to If, may exist in solvated as well as unsolvatedforms such as, for example, hydrated forms. It is to be understood thatthe invention encompasses all such solvated forms that are capable ofsaccharide recognition.

It is also to be understood that certain compounds of the Formula (I),and sub-formulae la to If, may exhibit polymorphism, and that theinvention encompasses all such forms that are capable of sacchariderecognition.

Compounds of the Formula (I), and sub-formulae la to Ig, may exist in anumber of different tautomeric forms and references to compounds of theFormula (I), and sub-formulae la to If, include all such forms. For theavoidance of doubt, where a compound can exist in one of severaltautomeric forms, and only one is specifically described or shown, allothers are nevertheless embraced by Formula I. Examples of tautomericforms include keto-, enol-, and enolate-forms, as in, for example, thefollowing tautomeric pairs: keto/enol (illustrated below),imine/enamine, amide/imino alcohol, amidine/amidine, nitroso/oxime,thioketone/enethiol, and nitro/aci-nitro.

Compounds of the Formula (I), and sub-formulae la to Ig, containing anamine function may also form N-oxides. A reference herein to a compoundof the Formula I that contains an amine function also includes theN-oxide. Where a compound contains several amine functions, one or morethan one nitrogen atom may be oxidised to form an N-oxide. Particularexamples of N-oxides are the N-oxides of a tertiary amine or a nitrogenatom of a nitrogen-containing heterocycle. N-Oxides can be formed bytreatment of the corresponding amine with an oxidizing agent such ashydrogen peroxide or a per-acid (e.g. a peroxycarboxylic acid), see forexample Advanced Organic Chemistry, by Jerry March, 4th Edition, WileyInterscience, pages. More particularly, N-oxides can be made by theprocedure of L. W. Deady (Syn. Comm. 1977, 7, 509-514) in which theamine compound is reacted with m-chloroperoxybenzoic acid (mCPBA), forexample, in an inert solvent such as dichloromethane.

Though the present invention may relate to any compound or particulargroup of compounds defined herein by way of optional, preferred orsuitable features or otherwise in terms of particular embodiments, thepresent invention may also relate to any compound or particular group ofcompounds that specifically excludes said optional, preferred orsuitable features or particular embodiments.

Suitably, the present invention excludes any individual compounds notpossessing the saccharide binding capabilites defined herein.

Immobilisation

In an embodiment, the compound of the present invention is immobilisedon or in a solid or semi-soild support. The person skilled in the art oforganic, synthetic chemistry will understand the term “solid orsemi-solid support” to refer to any suitable support in which thecompound of the invention may be immobilised on or incorporated within.Suitably, the solid and/or semi-solid support is selected from apolymeric matrix (e.g. a polystyrene bead) and/or a gel (e.g. a hydrogelor sol-gel). More suitably, the solid support is a polymeric matrix(e.g. a polystyrene bead) and the semi-solid support is a gel (e.g. ahydrogel or solgel).

In an embodiment, the polymeric matrix and/or gel comprises one or morehomopolymer, copolymer and/or crosslinked polymer. Suitably, thepolymeric matrix and/or gel comprises one or more polymer selected frompolyethylene glycol, poloxamer, polyacrylamide, polyacrylate,polyalkylacrylate, polyvinylpyrrolidine, polyvinylalcohol, polystyrene,polycarboxylate ethers, polyurethanes, polyallyamine, polyethylenimine,polysaccharides and mixtures and/or derivatives thereof.

In an embodiment, the polymeric matrix and/or gel comprises one or morewater soluble polymer. Suitably, the polymeric matrix and/or gelcomprises one or more water soluble polymer selected from polyethyleneglycol, polyacrylamide, polyvinylalcohol and polycarboxylate. Moresuitably, the polymeric matrix and/or gel comprises one or more watersoluble polymer selected from polyethylene glycol or polyacrylamide.

It will be understood that the compound of the present invention may beattached (immobilised) to the solid or semi-soild support by anysuitable means known in the art. The attachment of the compound of thepresent invention to the solid or semi-solid support may therefore takethe form of one or more covalent and/or non-covalent interactions.

In an embodiment, the compound of the present invention is chemicallylinked (covalently attached) to the polymeric matrix and/or gel. Thecompound of the present invention may be chemically linked to thepolymeric matrix and/or gel at any suitable position of the compound andthe attachment may take the form of any suitable bond. Suitably, thecompound of the present invention is chemically linked to the polymericmatrix and/or gel via one or more of the substitutent groups associatedwith at least one of the substituent groups R¹, R², R³, R⁴, R¹, Z¹, Z²,Z³, Z⁴ or Z⁵. More suitably, the the compound of the present inventionis chemically linked to the polymeric matrix and/or gel via one or moreof the substitutent groups associated with at least one of thesubstituent groups Z₁, Z₂, Z₃, Z₄ or Z₅.

In an embodiment, the compound of the present invention is chemicallylinked (covalently attached) to the polymeric matrix and/or gel via alinker, L₂. It will be understood that the linker, L₂, may be any groupcapable of forming a covalent attachment between the compound of thepresent invention and the polymeric matrix and/or gel. The linker, L₂,may take the form of a bond (e.g. an amide bond) or may be in the formof a suitable cross-linker molecule, used to couple together thecompound of the present invention and the polymeric matrix and/or gel.The person skilled in the art will be able to select suitablecross-linker molecules for use in covalently coupling the compounds ofthe present invention to the polymeric matrix and/or gel.

In another embodiment, the compound of the present invention isassociated with and/or physically incorporated within the polymericmatrix and/or gel via non-covalent interactions. It will be understoodthat any suitable non-covalent interaction may be utilised forassociation between the compound of the present invention and thepolymeric matrix and/or gel. Non-limiting examples of suitablenon-covalent interactions include: hydrogen-bonding interactions, ionicinteractions, hydrophobic interactions, van der Waal interactions andcombinations thereof.

Synthesis

The compounds of the present invention can be prepared by any suitabletechnique known in the art. Particular processes for the preparation ofthese compounds are described further in the accompanying examples.

In the description of the synthetic methods described herein and in anyreferenced synthetic methods that are used to prepare the startingmaterials, it is to be understood that all proposed reaction conditions,including choice of solvent, reaction atmosphere, reaction temperature,duration of the experiment and workup procedures, can be selected by aperson skilled in the art.

It is understood by one skilled in the art of organic synthesis that thefunctionality present on various portions of the molecule must becompatible with the reagents and reaction conditions utilised.

It will be appreciated that during the synthesis of the compounds of theinvention in the processes defined herein, or during the synthesis ofcertain starting materials, it may be desirable to protect certainsubstituent groups to prevent their undesired reaction. The skilledchemist will appreciate when such protection is required, and how suchprotecting groups may be put in place, and later removed.

For examples of protecting groups see one of the many general texts onthe subject, for example, ‘Protective Groups in Organic Synthesis’ byTheodora Green (publisher: John Wiley & Sons). Protecting groups may beremoved by any convenient method described in the literature or known tothe skilled chemist as appropriate for the removal of the protectinggroup in question, such methods being chosen so as to effect removal ofthe protecting group with the minimum disturbance of groups elsewhere inthe molecule.

Thus, if reactants include, for example, groups such as amino, carboxyor hydroxy it may be desirable to protect the group in some of thereactions mentioned herein.

By way of example, a suitable protecting group for an amino oralkylamino group is, for example, an acyl group, for example an alkanoylgroup such as acetyl, an alkoxycarbonyl group, for example amethoxycarbonyl, ethoxycarbonyl or t-butoxycarbonyl group, anarylmethoxycarbonyl group, for example benzyloxycarbonyl, or an aroylgroup, for example benzoyl. The deprotection conditions for the aboveprotecting groups necessarily vary with the choice of protecting group.Thus, for example, an acyl group such as an alkanoyl or alkoxycarbonylgroup or an aroyl group may be removed by, for example, hydrolysis witha suitable base such as an alkali metal hydroxide, for example lithiumor sodium hydroxide. Alternatively, an acyl group such as atert-butoxycarbonyl group may be removed, for example, by treatment witha suitable acid as hydrochloric, sulfuric or phosphoric acid ortrifluoroacetic acid and an arylmethoxycarbonyl group such as abenzyloxycarbonyl group may be removed, for example, by hydrogenationover a catalyst such as palladium-on-carbon, or by treatment with aLewis acid for example boron tris(trifluoroacetate). A suitablealternative protecting group for a primary amino group is, for example,a phthaloyl group which may be removed by treatment with an alkylamine,for example dimethylaminopropylamine, or with hydrazine.

A suitable protecting group for a hydroxy group is, for example, an acylgroup, for example an alkanoyl group such as acetyl, an aroyl group, forexample benzoyl, or an arylmethyl group, for example benzyl. Thedeprotection conditions for the above protecting groups will necessarilyvary with the choice of protecting group. Thus, for example, an acylgroup such as an alkanoyl or an aroyl group may be removed, for example,by hydrolysis with a suitable base such as an alkali metal hydroxide,for example lithium, sodium hydroxide or ammonia. Alternatively, anarylmethyl group such as a benzyl group may be removed, for example, byhydrogenation over a catalyst such as palladium-on-carbon.

A suitable protecting group for a carboxy group is, for example, anesterifying group, for example a methyl or an ethyl group which may beremoved, for example, by hydrolysis with a base such as sodiumhydroxide, or for example a t-butyl group which may be removed, forexample, by treatment with an acid, for example an organic acid such astrifluoroacetic acid, or for example a benzyl group which may beremoved, for example, by hydrogenation over a catalyst such aspalladium-on-carbon.

Resins may also be used as a protecting group.

The methodology employed to synthesise a compound of Formula (I) willvary depending on the nature of Rings A and B, C, D, R₁, R₂, R₃, R₄, W₁,W₂, W₃, W₄, X₁, X₂, X₃, X₄, Z₁, Z₂, Z₃, Z₄, Z₅, L, a, b, c, d, m, n, o,p and any substituent groups associated therewith. Suitable processesfor their preparation are described further in the accompanyingExamples.

In certain embodiments, the compounds of the present invention (i.e. thecompounds of Formula (I)) are prepared according to Method A or MethodB, shown below:

-   Method A    -   reacting a compound of Formula III, as shown below:

    -   

    -   wherein, bonds b₁ and b2, Rings A and B, D, W₁, W₄, X₁, X₄, Z₁,        Z₂, Z₄, R₁, R₂, R₄ and integers a, b, d, m, n and p are as        defined hereinabove, and q¹ is an integer selected from 0 or 1;

    -   with a compound of Formula IV:

    -   

    -   wherein, C, R₃, Z₃, c and o are as defined hereinabove, w¹ is        and integer from 0 to 1 and E₁, E₂ and E₃ are each selected from        a group of Formula X1, shown below:

    -   

    -   wherein:        -   

        -   denotes the point of attachment; and

        -   Y₁ is selected from O, S or NR_(j), wherein R_(j) is as            defined herein;

    -   and, optionally, thereafter, and if necessary:        -   i) removing any protecting groups present;        -   ii) converting the compound of Formula (I) into another            compound of Formula (I); and/or        -   iii) forming a salt, hydrate or solvate thereof;-   Method B    -   reacting a compound of Formula V, as shown below:

    -   

    -   wherein, bonds b₁ and b2, Rings A and B, D, W₁, W₄, X₁, X₄, Z₁,        Z₂, Z₄, R₁, R₂, R₄ and integers a, b, d, m, n and p are as        defined in hereinabove, q² is an integer selected from 0 or 1        and E₄, E₅ and E₆ are each selected from a group of Formula X2,        shown below:

    -   

    -   wherein:        -   

        -   denotes the point of attachment; and

        -   Y₂ is selected from O, S or NR_(j), wherein R_(j) is as            defined herein;

    -   with a compound of Formula VI:

    -   

    -   wherein, C, R₃, Z₃ and integers c and o are as defined in        hereinabove, and w² is and integer from 0 to 1;

    -   and, optionally, thereafter, and if necessary:        -   i) removing any protecting groups present;        -   ii) converting the compound of Formula (I) into another            compound of Formula (I); and/or        -   iii) forming a salt, hydrate or solvate thereof.

Suitably, Method A and/or B described hereinabove is carried out in thepresence of one or more of the following:

-   a base;-   a template-   a catalyst; and/or-   an activating agent.

In an embodiment, Method A and/or B is conducted in the presence of abase. Non-limiting examples of suitable bases include NaOH, KOH,potassium tert-butoxide, trimethylamine, diisopropylethylamine,diisopropylmethylamine, N-methylmorpholine, piperidine,2,2,6,6-tetramethylpiperidine, pyridine, 2,6-dimethylpyrridine,methylimidazole, 4-(Dimethylamino)pyridine (DMAP) and1,8-diazabicyclo(5.4.0)undec-7-ene (DBU). Suitably, the base ispyridine, 4-(Dimethylamino)pyridine (DMAP) or methylimidazole. Mostsuitably, the base is pyridine.

In another embodiment, Method A and/or Method B is conducted in thepresence of a template. The term “template” will be understood to be aterm of the art and refers to a molecule which is capable of reversiblyassociating with one or more of the starting materials and/orintermediates and/or final products of a reaction, and in doing so helpsto promote the generation of one or more of the final products of thereaction. Non-limiting examples of suitable templates includeoctyl-β-glucoside, methyl-β-glucoside, octyl-β-galactoside,methyl-β-galactoside, octyl-β-mannoside and methyl-β-mannoside.Suitably, the template is octyl-β-glucoside.

It will be understood that the template may be used in any suitableamount. Suitably, the mole ratio of template to the compound of FormulaIII or the compound of Formula V is from between 0.1:1 to 10:1. Moresuitably, the the mole ratio of template to the compound of Formula IIIor the compound of Formula V is from between 0.5:1 to 5:1. Mostsuitably, the mole ratio of template to the compound of Formula III orthe compound of Formula V is from between 0.5:1 to 2:1.

In a particular embodiment, Method A and/or Method B is conducted in thepresence of a base (e.g. 4-dimethylaminopyridine) and a template (e.g.octyl-β-glucoside).

In certain embodiments, Method A and B may be conducted in the presenceof one or both of a catalyst and/or an activating agent. The term“catalysts” will be understood to be any suitable reagent that helps topromote the rate of the reaction between the compounds of Formulae IIIand IV and V and VI without undergoing any permenant chemical change.Whereas, the term “activating agent” will be understood to be anysuitable agent that reacts with one or more of the starting materials ofthe reaction to help promote the reactivity of said starting material inthe reaction.

It will be appreciated that any suitable reaction conditions may be usedin Methods A and B defined hereinabove. Furthermore, it will beunderstood that the reaction conditions used in Methods A and B willvary according to the specific functional groups present. A personskilled in the art will be able to select suitable reaction conditions(e.g. temperature, pressures, reaction times, concentration etc.) to usein either Method A or Method B.

In an embodiment, Method A and/or B is conducted at a temperature ofbetween -100° C. and 200° C. Suitably, the process of the presentinvention is conducted at a temperature of between 0° C. and 150° C.More suitably, the process of the present invention is conducted at atemperature of between 0° C. and 100° C. Most suitably, the process ofthe present invention is conducted at a temperature of between 0° C. and75° C.

In another embodiment, Method A and/or B is carried out in an organicsolvent. The organic solvent may be used to solubilise the compounds ofFormulae III, IV, V and VI thereby facilitate reaction therebetween.Accordingly, it will be understood that the organic solvent selectedwill depend on the specific compound selected. Suitable organic solventsmay include, but are not limited to, chloroform, dichloromethane, DMF,DMSO, acetonitrile, tetrahydrofuran (THF), N-Methyl-2-pyrrolidone (NMP),2-methyltetrahydrofuran (2M-THF) and mixtures thereof.

In certain embodiments, Method A and/or Method B is carried out inpyridine.

In another embodiment, Method A and/or B is carried out under anhydrousconditions.

In further embodiment, Methods A and B are conducted under an inertatmosphere (i.e. under nitrogren or argon).

The resultant compounds of the present invention (i.e. compounds ofFormula (I)) may be isolated and purified using techniques well known inthe art. A non-limiting example of a suitable technique ischromatography, particularly high performance liquid chromatography(HPLC).

Intermediates

In another aspect, the present invention provides novel intermediates,as defined herein, which are suitable for use in the synthetic methodsas set out herein.

Thus, in a particular aspect of the present invention, there is provideda compound of Formula III or Formula V, or a salt, solvate, ester orhydrate thereof, as shown below:

wherein, each of bonds, b₁ and b2, Rings A and B, D, W₁, W₄, X₁, X₄, Z₁,Z₂, Z₄, R₁, R₂, R₄, a, b, d, m, n, p, E₄, E₅, E₆, q¹ and q² are asdefined hereinabove.

Preferred and suitable substituent groups for each of Rings A and B, D,W₁, W₄, X₁, X₄, Z₁, Z₂, Z₄, R₁, R₂, R₄, a, b, d, m, n and p in respectof the compounds of Formula III will be understood to be analogous tothe preferred and suitable substituent groups for each of Rings A and B,D, W₁, W₄, X₁, X₄, Z₁, Z₂, Z₄, R₁, R₂, R₄, a, b, d, m, n and p for thecompounds of Formula (I) described hereinabove.

In certain embodiments, the compounds of Formula III and/or Formula Vmay be optionally attached to a substituent group of Formula A1 asdefined hereinabove, at a position associated with one or more of thesubstituent groups R_(1a), R_(1b), R_(2a), R_(2b), R₁, R₂, R₃, R₄, Z₁,Z₂, Z₃, Z₄ and/or Z₅.

In a particular embodiment, the present invention provides the followingcompounds, or salts, solvates, ester or hydrates thereof:

wherein:

-   R^(z) is selected from -OCH₂C(O)ONa, -OCH₂C(O)OC(CH₃)₃ or    -OCH₂C(O)OCH₃;

-   R₁ is a group of the formula:

-   

-   each of Z₁, Z₂ and Z₅ is a group of the formula:

-   

-   each Z₁₀₀ is a group of the formula:

-   

-   and Z_(1a), Z_(2a) and Z_(5a) are selected from:

-   

-   -   wherein

    -   

    -   denotes the point of attachment; or

    -   

    -   wherein Z_(1′), Z_(2′) and Z_(5′) is a group of the formula:

    -   

In a particular embodiment, the present invention provides the followingcompounds, or salts, solvates, ester or hydrates thereof:

wherein:

-   R^(Z) is selected from —OCH₂C(O)ONa, —OCH₂C(O)OC(CH₃)₃ or    —OCH₂C(O)OCH₃; and each of Z₁, Z₂ and Z₅ is a group of the formula:

-   

-   -   wherein

    -   

    -   denotes the point of attachment; or

    -   

    -   wherein Z_(1′), Z_(2′) and Z_(5′) is a group of the formula:

    -   

In another particular embodiment, the present invention provides thefollowing compounds, or salts, solvates, ester or hydrates thereof:

wherein:

-   R^(z) is selected from —OCH₂C(O)ONa, —OCH₂C(O)OC(CH₃)₃ or    —OCH₂C(O)OCH₃; and each of Z₁, Z₂ and Z₅ is a group of the formula:

-   

-   wherein

-   

-   denotes the point of attachment.

Saccharide Recognition

The compounds of the present invention are advantageously capable ofassociation with one or more target saccharides in an aqueous medium. Itwill be understood that the association between the compounds of thepresent invention and the one or more target saccharides may involve oneor more covalent and/or non-covalent interactions therebetween.Suitably, the compounds of the present invention are capable ofassociation with the one or more target saccharides through non-covalentinterations (e.g. CH-Π interactions, van der Waal interactions and polarinteractions) only, which advantageously makes the compounds of thepresent invention suitable for use in reversible saccharide associationand subsequent continuous saccharide detection.

The isothermal titration calorimetry (ITC) studies described in theExamples section hereinbelow may be used to measure the saccharidebinding affinity of the compounds of the present invention. It will beappreciated that other suitable techniques known in the art may alsosimilarly be used to determine saccharide binding affinity. Non-limitingexamples of other suitable techniques include fluorescence titrations,UV-vis titrations and/or ¹H NMR titrations.

Although the saccharide binding affinities of the compounds of Formula Ivary with structural change, as expected, the compounds of the inventionwere found to demonstrate saccharide (e.g. glucose) binding affinity inITC studies described in the Examples section hereinbelow.

In general, the compounds of the invention demonstrate a bindingaffinity (K_(a)) towards the target saccharide (e.g. glucose) in waterof equal to or greater than 10 M⁻¹ in the ITC study described in theExamples section hereinbelow. Suitably, the the compounds of theinvention demonstrate a binding affinity (K_(a)) towards the targetsaccharide (e.g. glucose) in water of equal to or greater than 50 M⁻¹.More suitably the compounds of the invention demonstrate a bindingaffinity (K_(a)) towards the target saccharide (e.g. glucose) in waterof equal to or greater than 100 M⁻¹. Yet more suitably, the compounds ofthe invention demonstrate a binding affinity (K_(a)) towards the targetsaccharide (e.g. glucose) in water of equal to or greater than 500 M⁻¹.Even more suitably, the compounds of the invention demonstrate a bindingaffinity (K_(a)) towards the target saccharide (e.g. glucose) in waterof equal to or greater than 1000 M⁻¹. Most suitably, the compounds ofthe invention demonstrate a binding affinity (K_(a)) towards the targetsaccharide (e.g. glucose) in water of equal to or greater than 2000 M⁻¹.

In a particular embodiment, the compounds of the invention demonstrate abinding affinity (K_(a)) towards the target saccharide (e.g. glucose) inwater of equal to or greater than 5000 M⁻¹, most suitably, equal to orgreater than 10000 M⁻¹.

The compounds of the present invention also advantageously demonstrate aselectivity towards saccharides comprising all-equatorial substituentsover those comprising at least one axial substituent (e.g. mannose). Incertain embodiment, the compounds of the present invention alsodemonstrate a selectivity towards monosaccharides over di-, tri andoligosaccharides.

Suitably, the compounds of the present invention display a bindingaffinity (K_(a)) towards saccharides comprising all-equatorialsubstituents (e.g. glucose) which is at least 2 times higher than thebinding affinity (K_(a)) displayed towards saccharides comprising atleast one axial substituent (e.g. mannose). More suitably, the compoundsof the present invention display a binding affinity (K_(a)) towardssaccharides comprising all-equatorial substituents (e.g. glucose) whichis at least 5 times higher than the binding affinity (K_(a)) displayedtowards saccharides comprising at least one axial substituent (e.g.mannose). Yet more suitably, the compounds of the present inventiondisplay a binding affinity (K_(a)) towards saccharides comprisingall-equatorial substituents (e.g. glucose) which is at least 50 timeshigher than the binding affinity (K_(a)) displayed towards saccharidescomprising at least one axial substituent (e.g. mannose). Even moresuitably, the compounds of the present invention display a bindingaffinity (K_(a)) towards saccharides comprising all-equatorialsubstituents (e.g. glucose) which is at least 100 times higher than thebinding affinity (K_(a)) displayed towards saccharides comprising atleast one axial substituent (e.g. mannose). Most suitably, the compoundsof the present invention display a binding affinity (K_(a)) towardssaccharides comprising all-equatorial substituents (e.g. glucose) whichis at least 500 times higher than the binding affinity (K_(a)) displayedtowards saccharides comprising at least one axial substituent (e.g.mannose).

Furthermore, the compounds of the present invention also advantageouslydemonstrate a selectivity towards saccharides comprising all-equatorialsubstituents (e.g. glucose) over other commonly occurring smallmolecules. Non-limiting examples of such commonly occurring smallmolecules include, purines and pyrimidines (e.g. cytidine, adenosine,guanosine, uridine, adenine, cytosine, thymine, uracil, uric acid,hypoxantine and xanthine), organic acids (e.g. glutaric acid andglutamic acid) and amino acids (e.g. histidine, phenylanaline,tryptophan etc.). One problem commonly associated with saccharidereceptors known in the art is that they often display some affinitytowards such small molecules, which effectively ‘poisons’ the receptorsaffinity towards the target saccharide when such small molecules arepresent. Advantageously, compounds of the present invention show littleto no affinity towards such small molecules and thus do not suffer fromsuch ‘poisoning’ effects.

Complexes and Compositions

According to a further aspect of the invention, there is provided acomplex comprising a compound of the present invention, as definedherein, in association with a target saccharide.

It will be appreciated that preferred and suitable compounds of thepresent invention in relation to the complex will be analogous to thepreferred and suitable compounds described hereinabove in respect of thecompounds of the present invention per se.

In an embodiment, the target sacharride is a saccharide comprising allequatorial substituents. More suitably, the target saccharide is amonosaccharide comprising all equatorial substituents. Most suitably,the target saccharide is glucose (e.g. β-glucose).

In another aspect of the present invention, there is provided a complexcomprising a compound of the present invention, as defined herein, inassociation with a displaceable reporter molecule.

In an embodiment, the complex comprises a compound of Formula Ib, Ic orId, as defined herein, in association with a displaceable receptormolecule.

The displaceable reporter molecule will be understood to be any compoundwhich is capable of association (binding) with the compound of thepresent invention in the absence of a target saccharide (e.g. glucose),and which is capable of dissociation from the compound of the presentinvention, and detectable, of upon exposure of the composition to atarget saccharide.

In an embodiment, the displaceable reporter molecule is an aromaticmolecule and/or dye molecule. Suitably, the displaceable reportermolecule is an aromatic molecule. More suitably, the displaceablereporter molecule is a fluorescent aromatic molecule (e.g.fluoresceinamine or tetramethylrhodamine isothiocyanate).

Suitably, the displaceable reporter molecule has an emission wavelengthof between 300 nm and 1000 nm. More suitably, the displaceable reportermolecule has an emission wavelength of between 300 nm and 800 nm. Yetmore suitably, the displaceable reporter molecule has an emissionwavelength of between 500 nm and 700 nm.

In certain embodiments, the displaceable reporter molecule is attachedto a saccharide (e.g. a glucoside). Suitably, the displaceable reportermolecule is attached to a saccharide (e.g. a glucoside) via a linker(e.g. an alkyl linker).

According to a further aspect of the invention there is provided acomposition comprising a compound of the invention as definedhereinbefore, or a salt, hydrate or solvate thereof, and a displaceablereporter molecule which is capable of association with said compound.

In an embodiment, there is provided a composition comprising a compoundof Formula Ib, Ic or Id, as defined hereinabove, or a salt, hydrate orsolvate thereof, and a displaceable reporter molecule which is capableof association with said compound.

Suitably, the displaceable receptor molecule is an aromatic moleculeand/or dye molecule, more suitably an aromatic molecule, and mostsuitably a fluorescent aromatic molecule (e.g. fluoresceinamine ortetramethylrhodamine isothiocyanate).

In another embodiment, the composition of the invention comprises adiluent and/or carrier. Suitably, the diluent and/or carrier is apharmaceutically acceptable diluent and/or carrier, suitable for use in,for example, veterinary and/or medicinal applications (i.e.administration to an animal and/or human body).

According to a further aspect of the invention there is provided acomposition comprising a compound of the invention as definedhereinbefore, or a salt, hydrate or solvate thereof, and apharmaceutically acceptable diluent and/or carrier.

It will be appreciated that the compositions of the present inventionmay also comprise one or more additional excipients. Additionalexcipients may be included to improve various properties of theformulation, such as, for example, formulation stability,biocompatibility and administrability. A person skilled in the art willbe able to select suitable excipients to include based on conventionalknowledge in the formulation field.

A non-limiting list of possible additional excipients that may be addedto the compositions of the present invention include: pH modifiers,surfactants, viscosity modifiers, tonicity agents, sterilising agents,preservatives, lubricants and solubility enhancers.

In another embodiment, the composition of the present inventioncomprises one or more additional saccharide detection agents. Suitablesaccharide detection agents for incorporation in the composition of thepresent invention include known saccharide binding compounds (e.g.boronic acid based compounds) and/or saccharide specific enzymes (e.g.glucose oxidase (GOx)).

The composition of the invention may be obtained by any conventionalprocedure, using conventional formulation excipients, well known in theart.

APPLICATIONS, DEVICES AND KITS

The present invention provides compounds that display saccharide (e.g.glucose) binding affinity in water. Furthermore, in certain embodiments,the compounds of the present invention display a selectivity towardsall-equitorial saccharides over saccharides comprising one or more axialsubstituents.

The present invention therefore provides a use of a compound, as definedherein (e.g. a compound of Formula If or Ig), a complex, as definedherein, a composition, as defined herein, or a saccharide detectiondevice, as defined herein, for detecting a target saccharide in anaqueous environment. Suitably, the target saccharide is anall-equitorial saccharide, more suitably, an all-equitorialmonosaccharide, and most suitably, glucose (e.g. β-glucose).

In an embodiment, the aqueous environment is blood or blood plasma.

In another embodiment, the aqueous environment is a fermentation and/orcell culturing medium.

In embodiments in which a displaceable reporter molecule is used, itwill be appreciated that the displaceable reporter molecule isdissociated from the compound of the present invention, and subsequentlydetected, upon exposure of the composition, or saccharide detectiondevice, to a target saccharide (e.g. glucose).

According to another aspect of the present invention, there is provideda use of a complex, as defined herein, a composition, as defined herein,a saccharide detection device, as defined herein, or a compound, asdefined herein (e.g. a compound of Formula If or Ig), for the diagnosisof a condition which results in, or is otherwise associated with, anabnormal concentration of, and/or a change in the concentration of, atarget saccharide. Suitably, the target saccharide is an all-equitorialsaccharide, more suitably, an all-equitorial monosaccharide, and mostsuitably, glucose (e.g. β-glucose).

In an embodiment, the diagnosis of a condition which results in, or isotherwise associated with, an abnormal concentration of, and/or a changein the concentration of, a target saccharide is carried out in vivo.

In another embodiment, the diagnosis of a condition which results in, oris otherwise associated with, an abnormal concentration of, and/or achange in the concentration of, a target saccharide is carried out invitro or on a sample (i.e. blood sample) removed from the human and/oranimal body.

In a further embodiment, the condition is diabetes.

According to another aspect of the present invention there is provided asaccharide detection device comprising a composition, as defined herein,or a compound, as defined herein.

Suitably, the device is in a form compatible for introduction into ahuman and/or animal body. More suitably, the device is in a formcompatible for introduction into direct contact with the bloodstream ofa human and/or animal patient. Non-limiting examples of suitable devicesfor introduction into a human and/or animal body include a pellet,tablet, capsule, stent and/or chip.

In another embodiment, the device is in a form compatible forintroduction into a fermentation medium and/or cell culturing medium.Non-limiting examples of suitable devices for introduction into afermentation medium and/or cell culturing medium include a fibre opticcable and/or a stent.

The excellent binding affinities and selectivities displayed towardsglucose by certain compounds of the present invention makes themparticularly well suited for application in glucose responsive insulinbased systems. Glucose responsive insulin based systems are well knownby those skilled in the art, and are typically insulin based systemswhich are activated (e.g. switched on) by an increase in theconcentration of glucose.

Thus, in another aspect of the present invention, there is provided ause of a compound, as defined herein (e.g. a compound of Formula I, la,lb, Ic, Id, Ie, If or Ig), in a glucose responsive insulin based system.

According to another aspect of the present invention, there is provideda complex comprising a compound, as defined herein (e.g. a compound ofFormula I, la, Ib, Ic, Id, Ie, If or Ig) covalently attached to insulin.

According to yet another aspect of the present invention, there isprovided a kit comprising of a compound, as defined herein (e.g. acompound of Formula I, la, Ib, Ic, Id, le, If or Ig) and insulin.

In an embodiment, there is provided a kit comprising a compound ofFormula Ib, Ic or Id, as defined herein, and insulin.

According to a further aspect of the present invention, there isprovided a kit comprising a compound of the present invention and a(displaceable) reporter molecule.

In an embodiment, there is provided a kit comprising a compound ofFormula Ib, Ic or Id, as defined herein, and a (displaceable) reportermolecule.

EXAMPLES

Embodiments of the invention will be described, by way of example only,with reference to the accompanying drawings, in which:

FIG. 1 shows schematic illustations of the key interactions made betweenthe target saccharide and the compounds of the present invention (FIGS.1 a and 1 b ), as well as molecular models of a ground stateconformation of one particular compound of the present invention withglucose (FIGS. 1 c and 1 d ). In FIG. 1 c , ten intermolecular NH▪▪▪Ohydrogen bonds (with a distance of between 1.9 and 2.2 Å) can be seen,and FIG. 1 d further depicts the close CH-Π contacts made between thesaccharide and compound of the present invention.

FIG. 2 shows: a) the partial ¹H NMR spectra; and b) the binding analysiscurve for receptor 1 (0.25 mM) titrated with a combined solution ofD-glucose (9.6 mM) and receptor 1 (0.25 mM), in D₂O buffered with 10 mMphosphate buffer solution (pH 7.4) at 298 K. Spectra imply binding withslow exchange on NMR timescale. Integrations of peak at 8.04 ppm(denoted with •) versus region 8.22-7.21 ppm were plotted againstD-glucose concentration (mM). The calculated values for the integralsare overlaid with the observed values, giving K_(a) = 18,026 ± 208 M⁻¹(1.04%).

FIG. 3 shows the ¹H NMR spectra for receptor 1 (0.25 mM) titrated with acombined solution of D-glucose (9.6 mM) and receptor 1 (0.25 mM), in D₂Obuffered with 10 mM phosphate buffer solution (pH 7.4) at 298 K.

FIG. 4 shows the ¹H NMR spectra at certain time intervals of: a)α-D-glucose (5 mM, and b) α-D-glucose (5 mM) with receptor 1 (0.2 mM).Relative integrals of α-H1 (5.22 ppm) and β-H2 (3.23 ppm) protons overtime were calculated to determine if receptor 1 affected the rate ofanomerisation between α and β-D-glucose. Rate of anomerisation was foundto be independent of receptor 1 (see Table 2).

FIG. 5 shows a plot of relative integrals of αH1:βH2 versus time (min).Similar gradients suggest receptor does not affect the rate ofanomerisation of D-glucose.

FIG. 6 shows the ITC binding results for receptor 1 (0.13 mM) titratedwith glucose (7.5 mM) in H₂O, in which: A) shows the blank ITC run(addition of sugar into water); B) shows the actual run (sugar intoreceptor 1); C) shows the plotted change in enthalpy vs molar ratio andthe fit calculated with the supplied ITC software (K_(a) = 21,000 ± 2640M⁻¹); and D) shows the fit calculated using an Excel spreadsheet tocorroborate the result.

FIG. 7 shows the ITC binding results for receptor 1 (0.06 mM) titratedwith D-glucose (7 mM) in 10 mM PBS buffer (pH 7.4), in which: A) showsthe blank ITC run (addition of sugar into water); B) shows the actualrun (sugar into receptor 1); C) shows the plotted change in enthalpy vsmolar ratio and the fit calculated with the supplied ITC software (K_(a)= 19,100 ± 1310 M⁻¹); and D) shows the fit calculated using an Excelspreadsheet to corroborate the result.

FIG. 8 shows the ITC binding results for receptor 1 (0.06 mM) titratedwith D-glucose (7 mM) in 10 mM PBS buffer (pH 6), in which: A) shows theblank ITC run (addition of sugar into water); B) shows the actual run(sugar into receptor 1); C) shows the plotted change in enthalpy vsmolar ratio and the fit calculated with the supplied ITC software (K_(a)= 19,800 ± 1290 M⁻¹); and D) shows the fit calculated using an Excelspreadsheet to corroborate the result.

FIG. 9 shows the ITC binding results for receptor 1 (0.06 mM) titratedwith D-glucose (7 mM) in 10 mM PBS buffer (pH 8), in which: A) shows theblank ITC run (addition of sugar into water); B) shows the actual run(sugar into receptor 1); C) shows the plotted change in enthalpy vsmolar ratio and the fit calculated with the supplied ITC software (K_(a)= 23,400 ± 1850 M⁻¹); and D) shows the fit calculated using an Excelspreadsheet to corroborate the result.

FIG. 10 shows the ITC binding results for receptor 1 (0.06 mM) titratedwith D-glucose (7 mM) in DMEM Cell Culture Medium (no glucose, 10k MWCO,90% v/v) and 10 mM phosphate buffer solution (pH 7.4), in which: A)shows the blank ITC run (addition of sugar into medium); B) shows theactual run (sugar into receptor 1); C) shows the plotted change inenthalpy vs molar ratio; and D) shows the fit calculated using an Excelspreadsheet (Ka = 5637 ± 118 M⁻¹).

FIG. 11 shows the ITC binding results for receptor 1 (0.06 mM) titratedwith D-glucose (7 mM) in 10 mM phosphate buffer solution (pH 7.4) withadded salts: ferric nitrate (0.2 µM), calcium chloride (1.8 mM),magnesium sulfate (0.81 mM), potassium chloride (5.3 mM), sodiumbicarbonate (44 mM), sodium chloride (110 mM) and sodium phosphatemonobasic (0.9 mM), in which: A) shows the blank ITC run (addition ofsubstrate into medium); B) shows the actual run (substrate into receptor1); C) shows the plotted change in enthalpy vs molar ratio; and D) showsthe fit calculated using an Excel spreadsheet (Ka = 5164 ± 303 M⁻¹).

FIG. 12 shows ¹H NMR spectra showing receptor 1 (0.1 mM) dissolved inD₂O with 10 mM phosphate buffer (pH 7.4) at 298 K. Addition of MgSO₄(0.8 mM) and CaCl₂ (1.8 mM), which are the concentrations present inDMEM cell culture media, to free receptor showed a small change inchemical shift (δ in ppm) for proton s2. Addition of 2 equivalents (0.2mM) of D-glucose did not saturate the receptor. Addition of this sameconcentration of glucose to free receptor in D₂O with no added salts(top spectrum) did saturate the receptor, suggesting that Ca²⁺ and Mg²⁺inhibit binding.

FIG. 13 shows the ITC binding results for receptor 1 (0.06 mM) titratedwith D-glucose (7 mM) in Leibovitz’s L-15 Cell Culture Medium (noglucose, 10k MWCO, 90% v/v) and 10 mM phosphate buffer solution (pH7.4), in which: A) shows the blank ITC run (addition of sugar intomedium); B) shows the actual run (sugar into receptor 1); C) shows theplotted change in enthalpy vs molar ratio; and D) shows the fitcalculated using an Excel spreadsheet (Ka = 5214 ± 452 M⁻¹).

FIG. 14 shows the ITC binding results for receptor 1 (0.06 mM) titratedwith D-glucose (5 mM) in Human Blood Serum (no glucose, 10k MWCO, 90%v/v) and 10 mM phosphate buffer solution (pH 8.5), in which: A) showsthe blank ITC run (addition of sugar into medium); B) shows the actualrun (sugar into receptor 1); C) shows the plotted change in enthalpy vsmolar ratio; and D) shows the fit calculated using an Excel spreadsheet(Ka = 2477 ± 142 M⁻¹).

FIG. 15 shows: a) the ¹H NMR spectra; and b) the binding analysis curvefor receptor 1 (0.07 mM) titrated with a combined solution ofD-methyl-β-glucoside (10 mM) and receptor 1 (0.07 mM), in D₂O bufferedwith 10 mM phosphate buffer solution (pH 7.4) at 298 K. Spectra implybinding with slow exchange on NMR timescale. Integrations of peak at8.31 ppm (denoted with •) versus region 8.36-7.36 ppm were plottedagainst guest concentration (mM). The calculated values for theintegrals are overlaid with the observed values, giving K_(a) = 7522 ±414 M⁻¹ (5.51%).

FIG. 16 shows the ITC binding results for receptor 1 (0.13 mM) titratedwith methyl-β-D-glucoside (7 mM) in H₂O, in which: A) shows the blankITC run (addition of sugar into water); B) shows the actual run (sugarinto receptor 1); C) shows the plotted change in enthalpy vs molar ratioand the fit calculated with the supplied ITC software (K_(a) = 9120 ±542 M⁻¹); and D) shows the fit calculated using an Excel spreadsheet tocorroborate the result.

FIG. 17 shows the ITC binding results for receptor 1 (0.06 mM) titratedwith methyl-β-D-glucoside (7 mM) in 10 mM phosphate buffer solution (pH7.4), in which: A) shows the blank ITC run (addition of sugar intowater); B) shows the actual run (sugar into receptor 1); C) shows theplotted change in enthalpy vs molar ratio; and D) shows the fitcalculated using an Excel spreadsheet (K_(a) = 7886 ± 1296 M⁻¹).

FIG. 18 shows the ITC binding results for receptor 1 (0.1 mM) titratedwith D-glucuronic acid (5 mM) in 10 mM phosphate buffer solution (pH7.4) in which: A) shows the blank ITC run (addition of substrate intomedium); B) shows the actual run (substrate into receptor 1); C) showsthe plotted change in enthalpy vs molar ratio; and D) shows the fitcalculated using an Excel spreadsheet (Ka = 5348 ± 189 M⁻¹).

FIG. 19 shows the ¹H NMR spectra for receptor 1 (0.1 mM) titrated with acombined solution of D-gluconate (10 mM) and receptor 1 (0.1 mM), in D₂Obuffered with 10 mM phosphate buffer solution (pH 7.4) at 298 K. Spectraimply no binding was observed, despite some broadening of peaks at highconcentrations of guest.

FIG. 20 shows the ITC binding results for receptor 1 (0.06 mM) titratedwith Glucono-δ-lactone/gluconic acid (200 mM) in 10 mM phosphate buffersolution (pH 7.4), in which: A) shows the blank ITC run (addition ofsubstrate into water); B) shows the actual run (substrate into receptor1); C) shows the plotted change in enthalpy vs molar ratio.

FIG. 21 shows: a) the partial ¹H NMR spectra; and b) the bindinganalysis curve for receptor 1 (0.05 mM) titrated with a combinedsolution of D-galactose (250 mM) and receptor 1 (0.05 mM), in D₂Obuffered with 10 mM phosphate buffer solution (pH 7.4) at 298 K. Spectraimply binding with fast/intermediate exchange on NMR timescale. Changesin chemical shift (Δδ ppm) of peak at 7.63 ppm (denoted with •) wereplotted against increasing guest concentration (mM). The calculatedvalues for the Δδ are overlaid with the observed values giving K_(a) =132 ± 13 M⁻¹ (10.2%).

FIG. 22 shows for receptor 1 (0.06 mM) titrated with D-galactose (518mM) in H₂O, in which: A) shows the blank ITC run (addition of sugar intowater); B) shows the actual run (sugar into receptor 1); and C) showsthe plotted change in enthalpy vs molar ratio.

FIG. yre 23 shows the shows the ITC binding results for receptor 1 (0.1mM) titrated with D-galactose (75 mM) in 10 mM phosphate buffer solution(pH 7.4) in which: A) shows the blank ITC run (addition of substrateinto medium); B) shows the actual run (substrate into receptor 1); C)shows the plotted change in enthalpy vs molar ratio; and D) shows thefit calculated using an Excel spreadsheet (Ka = 182 ± 4.2 M⁻¹).

FIG. 24 shows the partial ¹H NMR spectra for receptor 1 (0.1 mM)titrated with a combined solution of 2-deoxy-D-glucose (50 mM) andreceptor 1 (0.1 mM), in D₂O buffered with 10 mM phosphate buffersolution (pH 7.4) at 298 K. Spectra imply binding with intermediate rateof exchange (rate between fast and slow exchange rates between H and HGspecies) on NMR timescale. Due to severe broadening of peaks forreceptor 1 upon addition of guest, no K_(a) was determinable.

FIG. 25 shows the ITC binding results for receptor 1 (0.06 mM) titratedwith 2-deoxy-D-glucose (7 mM) in H₂O, in which: A) shows the blank ITCrun (addition of sugar into water); B) shows the actual run (sugar intoreceptor 1); C) shows the plotted change in enthalpy vs molar ratio andthe fit calculated with the supplied ITC software (K_(a) = 657 ± 90M⁻¹); and D) shows the fit calculated using an Excel spreadsheet tocorroborate the result.

FIG. 26 shows the ITC binding results for receptor 1 (0.06 mM) titratedwith 2-deoxy-D-glucose (7 mM) in 10 mM phosphate buffer solution (pH7.4), in which: A) shows the blank ITC run (addition of sugar intowater); B) shows the actual run (sugar into receptor 1); C) shows theplotted change in enthalpy vs molar ratio; and D) shows the fitcalculated using an Excel spreadsheet (Ka = 725 ± 41 M⁻¹).

FIG. 27 shows: a) the partial ¹H NMR spectra; and b) the bindinganalysis curve for receptor 1 (0.11 mM) titrated with a combinedsolution of D-mannose (250 mM) and receptor 1 (0.11 mM), in D₂O bufferedwith 10 mM phosphate buffer solution (pH 7.4) at 298 K. Spectra implybinding with fast/intermediate exchange on NMR timescale. Changes inchemical shift (Δδ ppm) of peak at 7.63 ppm (denoted with •) wereplotted against increasing guest concentration (mM). The calculatedvalues for the Δδ are overlaid with the observed values giving K_(a) =140 ± 2M⁻¹ (1.31%).

FIG. 28 shows for receptor 1 (0.06 mM) titrated with D-Mannose (504 mM)in H₂O, in which: A) shows the blank ITC run (addition of sugar intowater); B) shows the actual run (sugar into receptor 1); and C) showsthe plotted change in enthalpy vs molar ratio.

FIG. 29 shows the ITC binding results for receptor 1 (0.1 mM) titratedwith D-mannose (75 mM) in 10 mM phosphate buffer solution (pH 7.4) inwhich: A) shows the blank ITC run (addition of substrate into medium);B) shows the actual run (substrate into receptor 1XX); C) shows theplotted change in enthalpy vs molar ratio; and D) shows the fitcalculated using an Excel spreadsheet (K_(a) = 143 ± 1.5 M⁻¹).

FIG. 30 shows the ITC binding results for receptor 1 (0.1 mM) titratedwith D-xylose (5 mM) in 10 mM phosphate buffer solution (pH 7.4) inwhich: A) shows the blank ITC run (addition of substrate into medium);B) shows the actual run (substrate into receptor 1); C) shows theplotted change in enthalpy vs molar ratio; and D) shows the fitcalculated using an Excel spreadsheet (K_(a) = 5804 ± 174 M⁻¹).

FIG. 31 shows the partial ¹H NMR spectra for receptor 1 (0.11 mM)titrated with a combined solution of D-cellobiose (250 mM) and receptor1 (0.11 mM), in D₂O buffered with 10 mM phosphate buffer solution (pH7.4) at 298 K. Spectra imply binding with slow exchange on NMRtimescale. Integrations of peak at 8.02 ppm (denoted with •) versusregion 8.36-7.36 ppm were used to calculate the K_(a) (M⁻¹) at eachpoint of addition (see Table 3), an average of these calculated valuesgives K_(a) = 31 ± 2.66 (9%).

FIG. 32 shows the ITC binding results for receptor 1 (0.06 mM) titratedwith cellobiose (250 mM) in H₂O, in which: A) shows the blank ITC run(addition of sugar into water); B) shows the actual run (sugar intoreceptor 1); C) shows the plotted change in enthalpy vs molar ratio andthe fit calculated with the supplied ITC software (K_(a) = 36.6 ± 2.5M⁻¹); and D) shows the fit calculated using an Excel spreadsheet tocorroborate the result

FIG. 33 shows the ITC binding results for receptor 1 (0.6 mM) titratedwith D-cellobiose (250 mM) in 10 mM phosphate buffer solution (pH 7.4)in which: A) shows the blank ITC run (addition of substrate intomedium); B) shows the actual run (substrate into receptor 1); C) showsthe plotted change in enthalpy vs molar ratio; and D) shows the fitcalculated using an Excel spreadsheet (K_(a) = 30.9 ± 4.9 M⁻¹).

FIG. 34 shows: a) the partial ¹H NMR spectra; and b) the bindinganalysis curve for receptor 1 (0.11 mM) titrated with a combinedsolution of D-fructose (250 mM) and receptor 1 (0.11 mM), in D₂Obuffered with 10 mM phosphate buffer solution (pH 7.4) at 298 K. Spectraimply binding with fast/intermediate exchange on NMR timescale. Changesin chemical shift (Δδ ppm) of peak at 7.63 ppm (denoted with •) wereplotted against increasing guest concentration (mM). The calculatedvalues for the Δδ are overlaid with the observed values giving K_(a) =51 ± 3 M⁻¹ (5.46%).

FIG. 35 shows the ITC binding results for receptor 1 (0.1 mM) titratedwith D-fructose (75 mM) in 10 mM phosphate buffer solution (pH 7.4) inwhich: A) shows the blank ITC run (addition of substrate into medium);B) shows the actual run (substrate into receptor 1); C) shows theplotted change in enthalpy vs molar ratio; and D) shows the fitcalculated using an Excel spreadsheet (K_(a) = 60.3 ± 1.6 M⁻¹).

FIG. 36 shows: a) the partial ¹H NMR spectra; and b) the bindinganalysis curve for receptor 1 (0.11 mM) titrated with a combinedsolution of D-ribose (250 mM) and receptor 1 (0.11 mM), in D₂O bufferedwith 10 mM phosphate buffer solution (pH 7.4) at 298 K. Spectra implybinding with fast exchange on NMR timescale. Changes in chemical shift(Δδ ppm) of peak at 7.83 ppm (denoted with •) were plotted againstincreasing guest concentration (mM). The calculated values for the Δδare overlaid with the observed values giving K_(a) = 264 ± 10 M⁻¹(3.96%).

FIG. 37 shows the ITC binding results for receptor 1 (0.1 mM) titratedwith D-ribose (75 mM) in 10 mM phosphate buffer solution (pH 7.4) inwhich: A) shows the blank ITC run (addition of substrate into medium);B) shows the actual run (substrate into receptor 1); C) shows theplotted change in enthalpy vs molar ratio; and D) shows the fitcalculated using an Excel spreadsheet (K_(a) = 216.5 ± 4.1 M⁻¹).

FIG. 38 shows: a) the partial ¹H NMR spectra; and b) the bindinganalysis curve for receptor 1 (0.1 mM) titrated with a combined solutionof methyl α-D-glucoside (500 mM) and receptor 1 (0.1 mM), in D₂Obuffered with 10 mM phosphate buffer solution (pH 7.4) at 298 K. Changesin chemical shift (Δδ ppm) of peak at 7.63 ppm (denoted with •) wereplotted against increasing guest concentration (mM). The calculatedvalues for the Δδ are overlaid with the observed values, which areeffectively indicative of no binding taking place.

FIG. 39 shows for receptor 1 (0.06 mM) titrated withmethyl-α-D-glucoside (500 mM) in H₂O, in which: A) shows the blank ITCrun (addition of sugar into water); B) shows the actual run (sugar intoreceptor 1); and C) shows the plotted change in enthalpy vs molar ratio.

FIG. 40 shows the ITC binding results for receptor 1 (0.06 mM) titratedwith methyl-α-D-glucoside (500 mM) in 10 mM phosphate buffer solution(pH 7.4), in which: A) shows the blank ITC run (addition of sugar intowater); B) shows the actual run (sugar into receptor 1) and C) shows theplotted change in enthalpy vs molar ratio.

FIG. 41 shows the ITC results for receptor 1 (0.06 mM) titrated withN-acetyl-D-glucosamine (498 mM) in H₂O, in which: A) shows the blank ITCrun (addition of sugar into water); B) shows the actual run (sugar intoreceptor 1); and C) shows the plotted change in enthalpy vs molar ratio.

FIG. 42 shows the ITC binding results for receptor 1 (0.06 mM) titratedwith N-acetyl-D-glucosamine (498 mM) in 10 mM phosphate buffer solution(pH 7.4), in which: A) shows the blank ITC run (addition of sugar intowater); B) shows the actual run (sugar into receptor 1) and C) shows theplotted change in enthalpy vs molar ratio.

FIG. 43 shows for receptor 1 (0.06 mM) titrated with D-uracil (5 mM) in10 mM PBS buffer (pH 7.4), in which: A) shows the blank ITC run(addition of sugar into water); B) shows the actual run (sugar intoreceptor 1); and C) shows the plotted change in enthalpy vs molar ratio.

FIG. 44 shows for receptor 1 (0.06 mM) titrated with uric acid (2.34 mM)in 10 mM PBS buffer (pH 7.4), in which: A) shows the blank ITC run(addition of sugar into water); B) shows the actual run (sugar intoreceptor 1); and C) shows the plotted change in enthalpy vs molar ratio.

FIG. 45 shows the ITC results for receptor 1 (0.06 mM) titrated withmaltose (500 mM) in H₂O, in which: A) shows the blank ITC run (additionof sugar into water); B) shows the actual run (sugar into receptor 1);and C) shows the plotted change in enthalpy vs molar ratio.

FIG. 46 shows the ITC binding results for receptor 1 (0.1 mM) titratedwith D-Mannitol (500 mM) in 10 mM phosphate buffer solution (pH 7.4) inwhich: A) shows the blank ITC run (addition of substrate into medium);B) shows the actual run (substrate into receptor 1); C) shows theplotted change in enthalpy vs molar ratio.

FIG. 47 shows the ITC binding results for receptor 1 (0.06 mM) titratedwith paracetamol (87 mM) in 10 mM phosphate buffer solution (pH 7.4), inwhich: A) shows the blank ITC run (addition of substrate into water); B)shows the actual run (substrate into receptor 1); C) shows the plottedchange in enthalpy vs molar ratio.

FIG. 48 shows the ITC binding results for receptor 1 (0.06 mM) titratedwith ascorbic acid (500 mM) in 10 mM phosphate buffer solution (pH 7.4),in which: A) shows the blank ITC run (addition of substrate into water);B) shows the actual run (substrate into receptor 1); C) shows theplotted change in enthalpy vs molar ratio.

FIG. 49 shows the ITC binding results for receptor 1 (0.06 mM) titratedwith L-fucose (500 mM) in 10 mM phosphate buffer solution (pH 7.4), inwhich: A) shows the blank ITC run (addition of substrate into water); B)shows the actual run (substrate into receptor 1); C) shows the plottedchange in enthalpy vs molar ratio.

FIG. 50 shows the ITC binding results for receptor 1 (0.06 mM) titratedwith L-phenylalanine (82 mM) in 10 mM phosphate buffer solution (pH7.4), in which: A) shows the blank ITC run (addition of substrate intomedium); B) shows the actual run (substrate into receptor 1); C) showsthe plotted change in enthalpy vs molar ratio.

FIG. 51 shows the ITC binding results for receptor 1 (0.1 mM) titratedwith myo inositol (5 mM) in 10 mM phosphate buffer solution (pH 7.4) inwhich: A) shows the blank ITC run (addition of substrate into medium);B) shows the actual run (substrate into receptor 1); C) shows theplotted change in enthalpy vs molar ratio; and D) shows the fitcalculated using an Excel spreadsheet (K_(a) = 7563 ± 313 M⁻¹).

FIG. 52 shows the ITC binding results for receptor 1 (0.1 mM) titratedwith Adenosine (500 mM) in 10 mM phosphate buffer solution (pH 7.4) inwhich: A) shows the blank ITC run (addition of substrate into medium);B) shows the actual run (substrate into receptor 1); C) shows theplotted change in enthalpy vs molar ratio.

FIG. 53 shows the ITC binding results for receptor 1 (0.1 mM) titratedwith cytosine (20 mM) in 10 mM phosphate buffer solution (pH 7.4) inwhich: A) shows the blank ITC run (addition of substrate into medium);B) shows the actual run (substrate into receptor 1); C) shows theplotted change in enthalpy vs molar ratio.

FIG. 54 shows the ITC binding results for receptor 1 (0.06 mM) titratedwith L-tryptophan (54 mM) in 10 mM phosphate buffer solution (pH 7.4),in which: A) shows the blank ITC run (addition of substrate intomedium); B) shows the actual run (substrate into receptor 1); C) showsthe plotted change in enthalpy vs molar ratio.

FIG. 55 shows the partial ¹H NMR ROESY spectrum of receptor 1 (2 mM)with D-glucose (5 mM, 2.5 equivalents) in D₂O. Chemical exchange peaks(black, annotated) link CH protons on β-D-glucose in free and boundstates. Chemical shifts for the glucose protons, with signal movementsdue to binding, are listed in the table. Signals for bound α-D-glucosewere not observed under these conditions.

FIG. 56 shows the structures of the substrates tested for affinity withreceptor 1.

FIG. 57 shows the partial ¹H NMR spectra (top) and binding analysiscurve (bottom) for 90 (1 mM) titrated with a combined solution ofD-glucose (1 M) and 90 (1 mM), in D₂O with at pH 7.4 and 298 K. Changein chemical shifts (Δδ, ppm) denoted with • were plotted againstD-glucose concentration (mM). The calculated values for Δδ are overlaidwith the observed values, giving K_(a) = 5.1 ± 0.2 M-1 (3.6%).

FIG. 58 shows the Partial ¹H NMR spectra (top) and binding analysiscurve (bottom) for 90 (0.25 mM) titrated with a combined solution ofD-cellobiose (250 mM) and 90 (0.25 mM), in D₂O with at pH 7.4 and 298 K.Change in chemical shifts (Δδ, ppm) denoted with • were plotted againstD-cellobiose concentration (mM). The calculated values for Δδ areoverlaid with the observed values, giving K_(a) = 46 ± 0.4 M-1 (0.89%).

FIG. 59 shows the partial ¹H NMR spectra (top) and binding analysiscurve (bottom) for 90 (0.2 mM) titrated with a combined solution ofD-cellotriose (15 mM) and 90 (0.2 mM), in D₂O with at pH 7.4 and 298 K.Change in chemical shifts (Δδ, ppm) denoted with • were plotted againstD-cellotriose concentration (mM). The calculated values for Δδ areoverlaid with the observed values, giving K_(a) = 949 ± 2.9 M-1 (0.3%).

FIG. 60 shows the partial ¹H NMR spectra for 90 (0.2 mM) titrated with acombined solution of D-cellotetraose (15 mM) and 90 (0.2 mM), in D₂Owith at pH 7.4 and 298 K. Spectra imply binding with intermediate rateof exchange, thus no K_(a) was determinable.

FIG. 61 shows the partial ¹H NMR spectra for 90 (0.2 mM) titrated with acombined solution of D-cellopentaose (15 mM) and 90 (0.2 mM), in D₂Owith at pH 7.4 and 298 K. Spectra imply binding with intermediate rateof exchange, thus no K_(a) was determinable.

FIG. 62 shows the partial ¹H NMR spectra (top) and binding analysiscurve (bottom) for 90 (0.2 mM) titrated with a combined solution ofD-maltose (500 mM) and 90 (0.2 mM), in D₂O with at pH 7.4 and 298 K.Change in chemical shifts (Δδ, ppm) denoted with • were plotted againstD-maltose concentration (mM). The calculated values for Δδ are overlaidwith the observed values, giving K_(a) = 15 ± 1.8 M-1 (11.8%).

FIG. 63 shows the partial ¹H NMR spectra (top) and binding analysiscurve (bottom) for 90 (0.2 mM) titrated with a combined solution ofD-maltotriose (500 mM) and 90 (0.2 mM), in D₂O with at pH 7.4 and 298 K.Change in chemical shifts (Δδ, ppm) denoted with • were plotted againstD-maltotriose concentration (mM). The calculated values for Δδ areoverlaid with the observed values, giving K_(a) = 20 ± 0.7 M-1 (3.3%).

FIG. 64 shows the ITC binding results for 90 (0.2 mM) titrated withD-cellobiose (200 mM) in water at 298 K, in which: A) shows the blankrun (addition of substrate into water); B) shows the titration(substrate into receptor); C) shows the plotted change in enthalpy vsmolar ratio; and D) shows the fit calculated using an Excel spreadsheet(K_(a) = 37.6 ± 2.5 M⁻¹).

FIG. 65 shows the ITC binding results for 90 (0.2 mM) titrated withD-cellotriose (15 mM) in water at 298 K, in which: A) shows the blankrun (addition of substrate into water); B) shows the titration(substrate into receptor); C) shows the plotted change in enthalpy vsmolar ratio; and D) shows the fit calculated using an Excel spreadsheet(K_(a) = 955 ± 11 M⁻¹).

FIG. 66 shows: a) the ITC titration of d-glucose (7.1 mM) in 10 mMphosphate buffer into Receptor 4 (0.40 mM) in 10 mM. at 298 K; and b) anenlarged image of the kcal mol⁻¹ of injectant vs molar ratio trace.K_(a) calculated at 6490 M-1 +/- 72.6 M⁻¹.

FIG. 67 shows: a) the ITC titration of d-glucose (7.1 mM) in 10 mMphosphate buffer into Receptor 5 (0.46 mM) in 10 mM. at 298 K; and b) anenlarged image of the kcal mol⁻¹ of injectant vs molar ratio trace.K_(a) calculated at 10400 M-1 +/- 132 M⁻¹.

FIG. 68 shows the ¹H NMR binding analysis curve generated following thetitration of a combined solution of β-D-glucose (10 mM) and Receptor 7(127 µM), in 10 mM PB, 140 mM NaCl, D₂O, into a solution of Receptor 7(127 µM) in 10 mM PB, 140 mM NaCl, D₂O. K_(a) calculated at 6886 M⁻¹ +/-190 M⁻¹.

FIG. 69 shows: a) the ITC titration of d-glucose (7.1 mM) in 10 mMphosphate buffer into Receptor 8 (0.42 mM) in 10 mM. at 298 K; and b) anenlarged image of the kcal mol⁻¹ of injectant vs molar ratio trace.K_(a) calculated at 4210 M⁻¹ +/- 73 M⁻¹.

FIG. 70 shows the ¹H NMR binding analysis curve generated following thetitration of a combined solution of β-D-glucose (10 mM) and Recepor 9(210 µM), in 10 mM PB, 140 mM NaCl, D₂O, into a solution of Receptor 9(210 µM) in 10 mM PB, 140 mM NaCl, D₂O.

FIG. 71 shows the ¹H NMR binding analysis curve generated following thetitration of a combined solution of β-D-glucose (100 mM) and Recepor 10(250 µM), in 10 mM PB, 140 mM NaCl, D₂O, into a solution of Receptor 10(250 µM) in 10 mM PB, 140 mM NaCl, D₂O.

FIG. 72 shows the ¹H NMR binding analysis curve generated following thetitration of a combined solution of β-D-glucose (10 mM) and Recepor 13(265 µM), in 10 mM PB, 140 mM NaCl, D₂O, into a solution of Receptor 13(265 µM) in 10 mM PB, 140 mM NaCl, D₂O.

FIG. 73 shows the the partial ¹H NMR spectra for Receptor 11 (50 µM) inD₂O (pH 7.4, 10 mM PBsoln) titrated with D-glucose (10 mM) with addedReceptor 11 (50 µM) and 10 mM PBsoln. In making the assumption ofreceptor saturation at ~1 mM, half saturation would be at 0.5 mM.Therefore 1/0.5 mM = K_(a) ~ 2000 M⁻¹.

FIG. 74 shows: A) the circular dichroism (CD) spectra; and B) thebinding analysis curve generated following the titration of D-glucose(10 mM) with added Receptor 11 (70 µM) and 10 mM PBsoln to a solution ofReceptor 11 (70 µM) in water (pH 7.4 with 10 mM PBsoln).

FIG. 75 shows: a) the ITC titration of d-glucose (7.73 mM) in 10 mMphosphate buffer into Receptor 13 (0.13 mM) in 10 mM at 298 K; and b) anenlarged image of the kcal mol⁻¹ of injectant vs molar ratio trace.K_(a) calculated at 1310 M⁻¹ +/- 33 M⁻¹.

FIG. 76 shows: a) the ITC titration of d-glucose (7.10 mM) in 10 mMphosphate buffer into Receptor 3 (0.29 mM) in 10 mM. at 298 K; and b) anenlarged image of the kcal mol⁻¹ of injectant vs molar ratio trace.K_(a) calculated at 5760 M⁻¹ +/- 269 M⁻¹.

FIG. 77 shows the ¹H NMR binding analysis curve generated following thetitration of a combined solution of β-D-glucose (3.24 M) and Recepor 2(265 µM) in D₂O, into a solution of Receptor 12 (223 µM) in D₂O at 298K.

Materials and Methods

Commercial reagents were purchased from Sigma-Aldrich, Alfa-Aesar orAcros Organics and were used without further purification unlessotherwise specified. All air and moisture sensitive manipulations werecarried out using standard vacuum line and Schlenk techniques, or in adrybox containing a purified argon atmosphere. Solvents for air andmoisture sensitive manipulations were obtained from an AnhydrousEngineering Solvent Purification System or distilled and dried overactivated molecular sieves.

Column chromatography was performed using silica gel 60 (Sigma Aldrich)and a suitable eluent. TLC was performed using aluminium backed TLCplates (Merck-Keiselgel 60 F254) and visualised using UV fluorescenceand/or developed using ninhydrin, potassium permanagante, EtOH/H₂SO₄,vanillin, Pd(OAc)₂/H₂O or iodine.

HPLC chromatography was performed using a Waters 600 Controller with aWaters 2998 Photodiode Array Detector. For analytical runs a XSELECT CSHC18 5 µm (4.6×150 mm) column was used and for preparative runs a XSELECTCSH Prep C18 5 µm OBD (19×250 mm) column was used, normally with anacetone-water solvent mixture.

¹H and ¹³C NMR spectra were recorded on Varian VNMR 400 MHz, JeolEclipse 400 MHz, Varian VNMR 500 MHz, Bruker cryogenically cooled 500MHz and Varian VNMR cryogenically cooled S600 MHz spectrometers. Allspectra were obtained at ambient temperature unless stated otherwise.All ¹H and ¹³C NMR chemical shifts are reported relative totetramethylsilane as an internal standard and in CDCl₃ unless otherwisestated, with ¹H (residual) and ¹³C chemical shifts of the solvent as asecondary standard.

IR spectra were recorded on Perkin-Elmer Spectrum One FT-IR spectrometerwith an ATR accessory and frequencies reported in wavenumbers (cm⁻¹).ESI-LRMS (electrospray ionisation low resolution mass spectrometry) wasperformed on a VG Analytical Quattro, ESI-HRMS (electrospray ionisationhigh resolution mass spectrometry) was performed on a Bruker DaltonicsApex IV and MALDI-MS (matrix-assisted laser desorption/ionisation) wasperformed on an Applied Biosystems 4700. Elemental analysis wasperformed on a EuroVector EA3000 Elemental Analyser.

¹H-NMR titrations were performed on a Varian VNMR cryogenically cooledS600 spectrometer. Solutions of saccharides in D₂O (99.9%), containingreceptor at a known concentration to be used in the experiment, wereprepared and allowed to equilibrate overnight before use if necessary.Aliquots were then added to an NMR tube containing a known concentrationof receptor solution (typically 100 µM - 400 µM). The receptorconcentration was therefore held constant while the carbohydrateconcentration was increased. The sample tube was shaken after eachaddition and ¹H-NMR spectra were acquired at 298 K.

Isothermal Titration (Micro)Calorimetry (ITC) experiments were performedon a MicroCal iTC200 microcalorimeter and/or a MicroCali VP-ITC. ITCexperiments were carried out at 298 K. Saccharide solutions wereprepared in HPLC-grade water and allowed to equilibrate overnight ifnecessary. The sample cell was charged with a known concentration ofreceptor solution in HPLC-grade water (typically 50 µM - 200 µM). Then,aliquots (typically 1.0 µL) of carbohydrate solution were added and theevolution of heat was followed as a function of time. Heats of dilutionwere measured by injecting the same carbohydrate solution intoHPLC-grade water, using identical conditions. For every addition, theheat of dilution was subtracted from the heat of binding using aMicroCal software programme implemented in ORIGIN 7.0. This gave an XYmatrix of heat vs. total guest concentration. This matrix was thenimported into a specially written Excel programme to fit the data to a1:1 binding model to give a Ka. ΔG can be derived from Ka, and ΔS can bederived from ΔH and ΔG using common thermodynamic equations. This methodof analysis was used in conjunction with fits for K_(a) calculated usingthe MicroCal software to corroborate the results obtained.

Synthetic Procedures Bicyclic Receptor Synthesis Scheme 1 — SyntheticProcedure Used to Prepare1,3,5-triethyl-2,4,6-tris(isocyanatomethyl)benzene (Compound 103)

1,3,5-Triethyl-2,4,6-tris(aminomethyl)benzene (Compound 106)

Under an inert N₂ atmosphere,1,3,5-tris(bromomethyl)-2,4,6-triethylbenzene 104 (324 mg, 0.74 mmol)was dissolved in anhydrous DMF (4.5 mL) and NaN₃ (157 mg, 2.42 mmol)added. The reaction was heated to 60° C. for 16 hours. The reactionmixture as then diluted with ethyl acetate (20 mL) and washed with water(3 × 20 mL), dried (MgSO₄) and filtered. DMF (4 mL) was added to thefiltrate and the solvent removed under vacuum to a volume of ~4 mL.Conversion to tris-azide 105 was confirmed by ¹H NMR (220 mg, 0.68 mmol,92%). The resultant DMF solution was transferred to a degassed anhydroussolution of THF (22 mL) and PMe₃ (1 M in THF, 4.1 mL) under an inert N₂atmosphere. The reaction mixture was stirred at room temperature for 1hour and degassed H₂O (5 mL) added, with the reaction mixture stirredfor a further 16 hours. The solvent and excess PMe₃ was then evaporatedby bubbling N₂ through the solution, and the crude residue suspended inH₂O (~10 mL). The suspension was then freeze dried to afford 106 (148mg, 0.61 mmol, 90%) as a white solid. ¹H NMR: (400 MHz, (CDCl₃): 1.24(t, J = 7.5 Hz, 9H, C(1)H), 2.83 (q, J = 7.5 Hz, 6H, C(2)H), 3.88 (s,6H, C(5)H₂); ¹³C NMR: (100 MHz, (CDCl₃): δ 16.8 (C(1)H₂), 22.6 (C(2)H),39.7 (G(5)H), 137.4 (C3), 140.4 (C4); LRMS: (ESI⁺) Found [M+Na]⁺: 272.2

1,3,5-Triethyl-2,4,6-tris(isocyanatomethyl)benzene (Compound 103)

Method A

A flask was charged with 106 (30 mg, 0.12 mmol) and NaHCO₃ (20 mg, 0.24mmol). CH₂Cl₂ (5 mL) and H₂O (5 mL) were added, the mixture cooled to 0°C. and rapidly stirred. Triphosgene (40 mg, 0.13 mmol) was added and thereaction mixture vigorously stirred at room temperature for 1 hour. Thereaction mixture was diluted with CH₂Cl₂ (20 mL) and brine (10 mL), andthe organic layer separated, dried (MgSO₄) and the solvent removed undervacuum to afford 103 (50 mg, 0.166 mmol, 78%) as a colourless oil.

Method B

Under an inert N₂ atmosphere, a flask was charged with triphosgene (1.81g, 6.1 mmol). Anhydrous toluene (70 mL) was added. A solution of 106(500 mg, 2.0 mmol) in anhydrous toluene (40 mL) was added dropwise overa period of 7 minutes. The reaction mixture was heated to reflux andstirred for a further 75 min. The reaction mixture was allowed to cooldown and the solvent removed under high vacuum. The residue was thenredissolved in about 40 mL toluene and filtered on cotton wool. Thesolvent was removed under high vacuum to afford 103 (630 mg, 1.9 mmol,95%) as an oil that slowly crystallised into a light yellow solid.

¹H NMR: (400 MHz, (CDCl₃): 1.26 (t, J = 7.6 Hz, 9H, C(1)H), 2.84 (q, J =7.6 Hz, 6H, C(2)H), 4.49 (s, 6H, C(5)H₂); ¹³C NMR: (100 MHz, (CDCl₃): δ16.1 (C(1)H₂), 22.8 (C(2)H), 40.4 (C(5)H), 123.1 (C6), 132.4 (C3), 143.1(C4); V _(max) 2973, 2933, 2875, 2243, 1495, 1453, 1335, 1042, 856, 577cm⁻¹; LRMS: (ESI⁺) Found [M+Na]⁺: 350.1. HRMS: (ESI⁺) calculated forC₁₈H₂₁N₃O₃Na⁺: 350.1475, found [M+Na]⁺: 350.1474.

Fmoc Protected Tert-butyl G2 Linker (Compound 84)

Method A

Under an inert N₂ atmosphere, Fmoc-amino benzoic acid 67 (983 mg, 2.63mmol), HBTU (996 mg, 2.63 mmol) and HOBt (355 mg, 2.63 mmol) weresuspended in anhydrous THF (30 mL). DIPEA (1.2 mL, 6.57 mmol) was addedand the reaction stirred at room temperature for 10 minutes. Secondgeneration dendritic amine 82 (3.1 g, 2.19 mmol) was then added and thereaction stirred for 24 hours. The solvent was then removed under vacuumand the crude residue purified by flash column chromatography (3%MeOH:CH₂Cl₂) to afford 84 (3.49 g, 2.01 mmol, 92%) as an off-whitesolid.

Method B

Under an inert N₂ atmosphere, 67 (5 g, 15.3 mmol), HBTU (5.03 g, 15.3mmol) and HOBt.H₂O (2.06 g mg, 15.3 mmol) were suspended in anhydrousTHF (60 mL). DIPEA (7.2 mL, 38.3 mmol) was added and the reactionstirred at room temperature for 1 hour. The solvent was removed undervacuum and the crude residue dissolved in ethyl acetate (100 mL) andthen poured into water (300 mL). The precipitate was then filtered andair dried to afford the crude HOBt ester/tetramethyl urea complex (1:1,4.36 g, 7.19 mmol, Mw = 607.67 g mol-¹) which was used without furtherpurification. The crude HOBt ester complex was then suspended inanhydrous THF (60 mL) and 82 (9.4 g, 6.54 mmol) and DIPEA (2.1 mL, 12.3mmol) added. The reaction mixture was stirred at room temperature for 24hours and the solvent removed under vacuum. The crude residue was thenpurified by flash column chromatography (5% MeOH:CH₂Cl₂) to afford 84(9.02 g, 5.19 mmol, 86%) as an off white solid.

¹H NMR: (400 MHz, (CDCl₃): δ 1.43 (s, 81H, C(26)H₃), 1.95 (m, 18H,C(23)H₂), 2.11 (t, J = 7.2 Hz, 6H, C(18)H₂), 2.17 (m, 18H, C(22)H₂),2.25 (t, J= 7.2 Hz, 6H, C(19)H₂), 4.27 (m, 3H, C(7)H and NH₂), 4.47 (d,J = 7.4 Hz, 2H, C(8)H₂), 6.08 (s, 3H, NH), 6.76 (d, J = 8.4 Hz, 1H,C(13)H), 7.26-7.31 (m, 2H, C(4)H), 7.38 (t, J = 7.4 Hz, 2H, C(3)H),7.57-7.69 (m, 2H, C(5)H), 7.71 (d, J= 8.7 Hz, 1H, C(2)H), 7.75 (d, J=7.6 Hz, 3H, C(2)H), 7.78 (d, J = 2.1 Hz, 1H, C(15)H), 8.54 (s, 1H, NH);¹³C NMR: (100 MHz, (CDCl₃): δ 28.0 (C26), 29.8 (C22), 29.9 (C23), 31.8(C19), 32.2 (C18), 47.2 (C7), 53.4 (C17), 57.4 (C21), 67.3 (C8), 80.6(C25) 116.6 (C12), 119.9 (C2), 122.6 (C10), 124.6 (C14), 125.3 (C4),126.0 (C15), 126.8 (C13), 127.0 (C5), 127.6 (C3), 141.3 (C1), 143.8(C6), 145.3 (C11), 154.9 (C9), 166.6 (C16), 172.7 (C24), 173.1 (C20); V_(max) 2977, 2963, 1752, 1723, 1689, 1637, 1535, 1367, 1242, 1151, 1098,844 cm⁻¹; HRMS: (ESI⁺) Found [M+2Na]²⁺ _(:) 921.0252.

Fmoc-amino benzoic acid (67) was prepared according to literatureprocedure as described in Angew. Chem., 2008, 120, 6957.

Second generation dendritic amine (82) was prepared according toliterature procedure as described in Angew. Chem. Int. Ed., 2015, 54,2057

Triamino G2MM Tert-butyl Protected Triethylbenzene Half Receptor(Compound 108)

Under an inert N₂ atmosphere, 84 (600 mg, 0.35 mmol) was dissolved in asolution of 103 (28 mg, 0.086 mmol) in anhydrous dichloromethane (5 mL).Anhydrous pyridine (40 µL) was added and the reaction heated to refluxfor 16 hours, after which it was cooled to room temperature and thesolvent removed under vacuum. The crude residue was purified by flashcolumn chromatography (1:1, EtOAc:CH₂Cl₂ ➔ 4% MeOH:CH₂Cl₂) to afford 107(400 mg, 0.072 mmol, 84%) as a white solid. Conversion to 107 wasconfirmed by limited NMR studies* and high resolution mass spectrometry(ESI⁺): m/z calculated for [M+3Na]³⁺ 2869.6877, found 1928.1169,calculated for [M+4Na]⁴⁺: 1451.8339, found 1451.8320. Under an inert N₂atmosphere, 107 (300 mg, 0.052 mmol) was dissolved in anhydrousdichloromethane (8 mL) and cooled to 0° C. DBU (50 µL, 0.30 mmol) wasadded dropwise and the reaction mixture warmed to room temperature andstirred for 1 hour. The solvent was removed under vacuum and the crudeproduct purified by flash column chromatography (4% MeOH:CH₂Cl₂) toafford 108 (238 mg, 0.047 mmol, 91%) as a white solid. ¹H NMR: (400 MHz,(CD₃OD): δ 1.24 (t, J = 7.6 Hz, C(1)H₃), 1.43 (s, 243H, C(23)H₃), 1.94(m, 54H, C(20)H₂), 2.09 (m, 18H, C(15)H₂), 2.18 (m, 54H, C(19)H₂), 2.23(m, 18H, C(16)H₂), 2.86 (m, 6H, C(2)H₂), 4.51 (s, 6H, C(5)H₂), 7.21 (dd,J = 2.1, 8.4 Hz, 3H, C(9)H), 7.29 (d, J = 2.1 Hz, 3H, C(11)H), 7.4 (d, J= 8.4 Hz, 3H, C(8)H), 7.4 (s, 9H, NH), 7.91 (s, 3H, NH); ¹³C NMR: (100MHz, (CDCl₃): δ 17.0 (C1), 22.5 (C2), 28.5 (C23), 30.5 (C19), 30.7(C20), 32.3 (C15), 32.6 (C16), 37.9 (C5), 58.9 (C18), 59.4 (C14), 81.6(C22), 117.4 (C11), 118.9 (C9) 124.5 (C8), 130.0 (C10), 132.9 (C3),133.8 (C7), 141.4 (C12), 145.1 (C4), 157.9 (C6), 170.1 (C13), 174.4(C21), 175.6 (C17); HRMS: (ESI⁺) Found [M+3Na]³⁺: 1706.0490.

* Limited NMR studies were only possible due to slow conformationalexchange of 107 resulting in very broad signals of low intensity.

Tert-butyl Protected Triethylbenzene Receptor (Compound 1a)

Method A

Under an inert N₂ atmosphere, 108 (124 mg, 0.024 mmol) was dissolved inanhydrous dichloromethane (50 ml) and heated to reflux. A solution of103 (8 mg, 0.024 mmol) in anhydrous dichloromethane (3 mL) was added andthe reaction stirred at reflux for 3 days. The reaction mixture wascooled to room temperature and the solvent removed under vacuum. Thecrude product was purified by reverse-phase HPLC and then freeze driedto afford 1a (20 mg, 0.004 mmol, 15%) as a white solid.

Method B

Under an inert N₂ atmosphere, 108 (200 mg, 0.04 mmol), Octyl-glucoside(23 mg, 0.08 mmol) and 4-dimethylaminopyridine (14 mg, 0.12 mmol) weredissolved in anhydrous dichloromethane (35 mL). A solution of 103 (13mg, 0.04 mmol) in anhydrous dichloromethane (5 mL) was added and thereaction stirred at reflux for 2 days. The reaction mixture as cooled toroom temperature and the solvent removed under vacuum. The crude productwas purified by reverse phase HPLC and then freeze dried to afford 1a(85 mg, 0.016 mmol, 40%) as a white solid.

¹H NMR: (400 MHz, (CD₃OD): δ 1.24 (m, 18H C(1)H₃), 1.43 (s, 243H,C(23)H₃), 1.95 (m, 54H, C(20)H₂), 2.13 (m, 18H, C(15)H₂), 2.20 (m, 54H,C(19)H₂), 2.25 (m, 18H, C(16)H₂), 2.74, 2.84 (br m, 6H, C(2)H₂), 4.40,4.49 (br s, 6H, C(5)H₂), 7.43 (s, 9H, NH), 7.63 (d, J = 8.7 Hz, 3H,C(10)H), 8.03 (d, J = 8.7 Hz, 3H, C(11)H), 8.07 (s, 3H, C(8)E); ¹³C NMR:(100 MHz, (CDCl₃): δ 15.5, 15.6 (C1), 22.3, 22.5 (C2), 27.1 (C23), 29.1(C19), 29.3 (C20), 30.8 (C15), 31.0 (C16), 37.5 (C5), 57.3 (C18), 58.1(C14), 80.2 (C22), 120.3 (C11), 123.5 (C8), 124.1 (C10), 127.6 (C7),129.4 (C9), 132.0, 132.6 (C3), 135.0 (C12), 143.0, 143.2 (C4), 155.8,156.6 (C6), 168.2 (C13), 173.0 (C21), 174.1 (C17); HRMS: (ESI⁺) Found[M+3Na]³⁺: 1816.1093.

Triethylbenzene Receptor (Receptor 1)

1a (3.5 mg, 0.65 µmol) was dissolved in HPLC grade dichloromethane (1.6ml). Trifluoroacetic acid (0.4 mL) was added and the reaction stirred atroom temperature for 16 hours. The solvent was then removed under a flowof nitrogen, the crude product suspended in water and freeze dried. Theresultant solid was then suspended in water, neutralised to pH 7 withNaOH (aq) and then freeze dried to afford receptor 1 (3.3 mg, 0.62 µmol,95%) as a white solid. ¹H NMR: (400 MHz, (D₂O): δ 1.17 (m, 18H C(1)H₃),1.94 (m, 54H, C(20)H₂), 2.12 (m, 18H, C(15)H₂), 2.18 (m, 54H, C(19)H₂),2.31 (m, 18H, C(16)H₂), 2.76 (br m, 6H, C(2)H₂), 4.46 (br s, 6H,C(5)H₂), 7.61 (d, J= 9.6 Hz, 3H, C(10)H), 7.82 (br m, 6H, C(11)H,C(8)H); ); ¹³C NMR: (100 MHz, (CDCl₃): δ 18.0, 18.1 (C1), 25.0, 25.2(C2), 32.6 (C15), 32.7 (C19), 32.9 (C16), 33.3 (C20), 40.1, 40.3 (C5),60.7 (C18), 61.3 (C14),120.0 (C11), 121.6 (C10), 122.8 (C8), 125.7 (C7),130.0 (C9), 134.7, 135.0 (C3), 135.6 (C12), 146.7, 146.9 (C4), 159.6,159.9 (C6), 170.3 (C13), 177.8 (C17), 184.1 (C17).

Compound 8d

A schlenk flask equipped with a magnetic stirrer was charged with 108(200.0 mg, 0.04 mmol), DMAP (14.5 mg, 0.12 mmol) and n-octyl glucoside(23.2 mg, 0.08 mmol) were added to the flask and placed under nitrogen.Anhydrous DCM (40 mL) was added and the reaction was warmed to 34° C. Asolution of compound 5d (12.5 mg, 0.051 mmol) in toluene (ca. 85%purity) was added to the flask and the reaction was allowed to stir for16 hours. The solvent was removed completely and the crude product waspurified by reverse phase MPLC on a C18 SNAP Ultra 60 g cartridgeeluting (10% acetone:water for 1 CV, a gradient of 10-95% acetone:waterfor 10 CV and 95% acetone:water for 2 CV). White solid (67 mg, 0.012mmol, 32 %).

Compound 8e - Receptor 2

Method A: Compound 8d was dissolved in anhydrous DCM (Vol: 6 mL) and TFA(1.74 mL, 22.67 mmol) at RT. The resulting off yellow solution wasstirred for 2 h at room temperature. Volatiles were removed under vacuumto give a yellow solid. The solid which was purified by reverse phaseMPLC on a C18 SNAP Ultra 60 g cartridge by loading the sample in 1:1MeOH/H₂O+0.1% formic acid). The resulting white solid was neutralisedusing 100 mM NaOH solution to pH 7 and the resulting solutionconcentrated to dryness under vacuum. White crystalline solid (37 mg,0.008 mmol, 74 %).

Method B: The solid obtained from Compound 44 was dissolved in anhydrousCH₂Cl₂ (5.9 mL) and TFA (1.7 mL, 23 mmol) was added at RT. The resultingoff yellow solution was stirred for 2 h at RT or until complete by TLC(product on the baseline in 60% EtOAc/CH₂Cl₂). The solvent/TFA wereremoved under vacuum (rotary evaporator then high vacuum) to give ayellow solid. The solid was purified by reverse phase chromatography(loading sample in 1:1 MeOH/H₂O+0.1 % formic acid). Fr 2-7 taken andconcentrated under vacuum. The resulting white solid was neutralisedusing 100 mM NaOH solution to pH 7 and the resulting solutionconcentrated under vacuum (rotary evaporator/cold finger with liquid N₂)to give Compound 45 as a white crystalline solid (37 mg, 75%).

¹H NMR: (500 MHz, D₂O, 298 K): δ 7.74 (br d, J = 7.1 Hz, 3H, H₉), 7.59(d, J = 8.6 Hz, 3H, H₈), 7.48 (s, 3H, NH), 7.40 (s, 3H, H₆), 7.01 (s,3H, H₁), 4.40 (br s, 6H, H₁₂), 3.93 (br s, 6H, H₃), 2.70 (br s, 6H,H₁₅), 2.20 (br s, 18H, H₂₀), 2.07 (m, 72H, H₁₉+H₂₄), 1.83 (m, 54H, H₂₃),1.07 (br t, 9H, H₁₆).

¹³C NMR (126 MHz, D₂O) δ 173.2, 166.0, 160.0, 149.4, 148.1, 135.5,131.1, 128.9, 122.8, 121.6, 119.6, 118.0, 117.4, 114.5, 113.9, 49.5,49.0, 48.9, 43.1, 33.8, 28.6, 22.0, 21.6, 21.2, 21.1, 13.5, 6.3.

Compound 9 - Receptor 3

Triethylbenzene receptor 1 (15.0 mg, 0.003 mol), HBTU (W: 2.6 mg, 0.006mmol) and 5-(aminoacetamido)fluorescein (21.8 mg, 0.048 mmol) weredissolved in DMF (1.25 mL) and H₂O (1.25 mL). The reaction mixture wasstirred in the dark, at room temperature for 16 hr then concentratedunder reduced pressure. The crude reaction mixture was neutralised to~pH 7 with aq. NaOH then concentrated under reduced pressure. Purifiedby reverse phase flash chromatography on a 12 g SNAP Ultra C18 cartridgeelution with 1CV 20% MeOH/H₂O +0.1% formic acid to 65% MeOH/H₂O +0.1%formic acid over 8 CV to 100% H₂O +0.1% formic acid over 0.5CV then 100%H₂O +0.1% formic acid for 3 CV. Product containing fractions werecombined, concentrated under reduced pressure, neutralised to ~pH 7 thenlyophilised to give compound 9 as an orange powder. ¹H NMR: (400 MHz,D₂O, 298 K): δ 7.91 - 7.28 (m, 15H, Ar), 7.25 - 6.89 (m, 9H, Ar), 6.73 -6.32 (m, 12H, Ar), 4.52 - 4.04 (m, 12H, H₅), 4.02 - 3.81 (m, 6H, H₂₂),2.59 - 1.61 (m, 156H, H₁₊₁₅₊₁₆₊₁₉₊₂₀), 1.21 - 0.82 (m, 18H, H₁).

Key Intermediates Compound A1

3,4-Diaminobenzoic acid (41.000 g, 0.269 mol) was mixed with SaturatedNaHCO₃ (0.40 L) and acetonitrile (0.40 L) to give a brown slurry. Next,solid Fmoc- OSu (99.99 g, 0.296 mol) was added in portions over 5minutes. The heterogenous suspension was allowed to stir at roomtemperature for 16 hours and then acidified with 1 M HCl(_(aq)). Thesolids were collected on a frit and washed with cold diethy ether (3×100 mL), hexane (3 × 100 mL), then MeOH (3 × 50 mL) and then driedunder vacuum. Brown solid (101 g, 0.269 mol, 100 %). This intermediate(10.000 g, 0.027 mol), HOBt (8.181 g, 0.053 mol), and HBTU (20.259 g,0.053 mol) were dissolved in THF (300) mL) and DIPEA (18.610 mL, 0.107mol). The heterogenous slurry was stirred at room temperature for 90minutes after which the solvent was removed in vacuo to afford a viscousoil. The oil was dissolved in EtOAc (80 ml) and was added to a rapidlystirring mixture of water (200 ml) and EtOAc (40 mL). After ca. 2 mins aprecipitate formed and diethyl ether (100 mL) was added to the flask.After stirring for 10 minutes, the solids were collected by filtrationand washed with water (3 × 10 mL) and diethyl ether (2 × 10 mL) beforedrying under vacuum for 16 hours. ¹H NMR: (500 MHz, DMSO-d₆) δ 8.94 (s,1H), 8.14 (dd, J = 17.7, 8.4 Hz, 2H), 8.01 - 7.93 (m, 1H), 7.90 (d, J =7.6 Hz, 2H), 7.87 - 7.68 (m, 6H), 7.68 - 7.61 (m, 1H), 7.54 (dt, J =11.5, 7.5 Hz, 1H), 7.42 (t, J = 7.4 Hz, 2H), 7.34 (t, J = 7.6 Hz, 2H),6.89 (d, J = 8.6 Hz, 1H), 6.46 (s, 2H), 4.44 (s, 2H), 4.31 (s, 1H), 3.40(s, 7H), 3.03 (s, 7H), 2.50 (s, 4H).

Compound 2

A 100 mL Schlenk flask was charged with compound A1 (2.750 g mg, 2.263mmol), N,N-diisopropylethylamine (0.585 mL, 3.341 mmol) and anhydrousTHF (10 mL). Solid 3-(2-aminoethyl)pentane-1,3,5-N-Boc-triamine* (0.800g, Moles: 1.911 mmol) was added and the homogenous solution/suspensionwas left to stir . After 72 hours, TLC (SiO₂, 7:3 EtOAc: DCM) indicatedcomplete consumption of starting materials and formation of the product.The solvent was removed in vacuo and the crude residue was extractedwith DCM (2 × 10 mL). The filtrate was concentrated to dryness and theresidue purified by MPLC (12%➔ 100%DCM in EtOAc). Orange amorphous solid(1.154 g, 1.413 mmol, 74 %). ¹H NMR: (400 Hz, CDCl₃) δ 7.00- 7.82 (m,11H, ArH), 6.71 (2H, d, J= 8.13 Hz, NH₂), 3.14 (6H, q, J= 7 Hz, CH₂NH),2.04 (app. s, 6H, CCH₂), 1.40 (27H, s, C(CH₃)₃). MS: (ESI⁺) calculatedfor C₄₄H₆₀N₆O₉Na⁺: 839.4314, found [M+Na]⁺: 839.4318.

*Carter, T. S.; Mooibroek, T. J.; Stewart, P. F. N.; Crump, M. P.;Galan, M. C. & Davis, A. P. Angewandte Chemie International Edition,2016, 55, 9311-9315.

Compound 3b

NaH (1.05 g, 26.3 mmol, 60% in mineral oil) was added to a Schlenk flask(100 mL) and placed under nitrogen. The mineral oil was removed bywashing the solids with 3× 25 mL petroleum ether 60-80° C. The washedNaH was suspended in anhydrous DMF (10 mL) and vigorously stirred forca. 10 mins whilst cooling in an ice bath. Solid trifluoroacetamide(4.46 g, 39.46 mmol) was added portion-wise under a nitrogencounter-flow. After five minutes of stirring the mixture was let warm toroom temperature. Once the evolution of gas had completely stopped(within 1 hour), solid 1,3,5-tribromomethyl-2,4,6-trimethoxylbenzene*(2.00 g, 4.39 mmol) was added portion-wise under a nitrogen counter-flowand the resulting white suspension was stirred at room temperature.After ca. 18 hours, the suspension was poured into 0.5 M HCl (150 mL)and the light orange precipitate collected on a frit. The solids werewashed with water (2 × 10 mL) and then dried under vacuum overnight (ca.10⁻² mbar). Off-white solid (2.44 g, 4.491 mol, 102 %). The crudeproduct was used without further purification and contained 2-3 % DMF(quantified by ¹H NMR spectroscopy). ¹H NMR: (400 MHz, DMSO-d₆) δ d.4.40 (6 H, ³J_(HH) = 4.6 Hz, ArCH₂NHC(O)CF₃), s. 3.70 (9 H, OCH₃). ¹³CNMR: (400 MHz, DMSO-d₆) δ q. 158.3 (²J_(CF) = 37.1 Hz, C(O)CF₃), 137.3,132.1 (Ar), q. 116.1 (¹J_(CF) = 287.9 Hz, CF₃), 62.5 (NHCH₂Ar), 32.1(OCH₃).

*Rosien, J.; Seichter, W.; Mazik, M., Organic and BiomolecularChemistry, 2013, 11(38), 6569-6579,

Compound 3c

Prepared by an analogous route using1,3,5-tribromomethyl-2,4,6-trimethylbenzene to Compound 3b. Off-whitesolid (4.86 g, 0.01 mol 82 %). ¹H NMR: (500 MHz, CDCl₃/MeOH-d₄) δ s.4.47 (6 H, ArCH₂NHC(O)CF₃), s. 2.26 (9 H, OCH₃). ¹³C NMR: (125 MHz,CDCl₃/MeOH-d₄) δ q. 157.6 (²J_(CF) = 37.2 Hz, C(O)CF₃), 137.8, 131.1(Ar), q. 115.9 (¹J_(CF) = 287.3 Hz, CF₃), 39.4 ArCH₂NHC(O)CF₃), 16.0(ArCH₃). MS :(APCl⁻) 493.9

Compound 3d

Prepared by an analogous route to Compound 3b using1,3,5-tris(bromomethyl)benzene (5.07 g, 0.014 mol). The crude productwas purified by reverse phase MPLC on a C18 SNAP Ultra 60 g cartridgeeluting (10% acetone:water for 1 CV, a gradient of 10-95% acetone:waterfor 10 CV and 95% acetone:water for 2 CV). White solid (2.16 g, 0.005mol, 34 %). ¹H NMR: (400 MHz, MeOH-d₄) δ 7.07, (s, 3H, ArH), 4.35 (6 H,³J_(HH) = 4.6 Hz, ArCH₂N(O)HCF₃). ¹³C NMR: (125 MHz, MeOH-d₄) δ q. 158.1(²J_(CF) = 36.9 Hz, C(O)CF₃), 138.1, 132.2 (Ar), q. 114.6 (¹J_(CF) =286.9 Hz, CF₃),125.9 (Ar), 42.3, ArCH₂NHC(O)CF₃).

Compound 4a

NaH (3.809 g, 0.095 mol, 60% in mineral oil) was added to a Schlenkflask (100 mL) and placed under nitrogen. The mineral oil was removed bywashing the solids with 3× 25 mL petroleum ether 60-80° C. The washedNaH was suspended in anhydrous DMF (40 mL) and vigorously stirred forca. 10 mins whilst cooling in an ice bath. Solid trifluoroacetamide(16.147 g, 0.143 mol) was added portion-wise under a nitrogencounter-flow. After five minutes of stirring the mixture was let warm toroom temperature. Once the evolution of gas had completely stopped(within 1 hour), solid 1,3,5-tribromomethyl-2,4,6-trimethoxylbenzene*(7.00 g, 0.016 mol) was added portion-wise under a nitrogen counter-flowand the resulting white suspension was stirred at room temperature.After ca. 18 hours, the suspension was poured into 0.5 M HCl (150 mL)and the light orange precipitate collected on a frit. The solids werewashed with water (2 × 10 mL) and then dried under vacuum overnight (ca.10⁻² mbar). Off-white solid (7.910 g, 0.015 mol 93 %). The intermediateacetamide (4.90 g, 0.009 mol) was dissolved in methanol (38.6 mL) andwater (38.6 mL). NaOH (1.05 g, 3.150 mol) was added and the reactionmixture was left to stir for ca. 18 hours at 65° C. Solid Boc₂O (7.287g, 0.033 mol) and triethylamine (2.534 mL, 0.026 mol) were added and thereaction was left to stir for a further 4 hours at ambient temperature.The reaction mixture was diluted with DCM (200 mL) and washed with sat.aq. NaHCO₃ (200 mL), 1 _(M) NaOH (200 mL) and brine (100 mL). Theorganic phase was concentrated to dryness and the resultant crudeproduct purified by MPLC (0➔50% MeOH in DCM). Colourless solid (4.820 g,0.009 mol, 96 %). ¹H NMR: (400 MHz, CDCl₃) δ m.br 4.33 (9 H,ArCH₂NHCO₂C(CH₃)₃), NH), q. 2.71 (6H, ³J_(HH) = 7.5 Hz, ArCH₂CH₃), s.1.44 (27H, CO₂C(CH₃)₃), t. 1.19 (9H, ³J_(HH) = 7.5 Hz, ArCH₂CH₃). ¹³CNMR: (125 MHz, CDCl₃) δ 155.5 (CO₂C(CH₃)₃), 143.9, 132.6 (Ar), 79.7(CO₂C(CH₃)₃), 38.9 ArCH₂NHCO₂C(CH₃)₃), 28.6 (CO₂C(CH₃)₃), 23.0(ArCH₂CH₃), 16.7 (ArCH₂CH₃).

*Rosien, J.; Seichter, W.; Mazik, M., Organic and BiomolecularChemistry, 2013, 11(38), 6569-6579.

Compound 4b

Compound 3c (9.123 g, 0.013 mol) was dissolved in MeOH (125 mL), andNaOH (1.689 g, 0.042 mol) at room temp overnight. 0.040 mol) was addedfollowed by BOC anhydride (11.702 g, 0.054 mol) and left to stirovernight. The solvent was removed under vacuum and crude productpartitioned between DCM and H₂O. The organic fractions were combined andwashed with 0.5 M HCl (50 ml), dried with MgSO₄ and concentrated undervacuum to give an off white solid. Purified by MPLC (0➔20% EtOAc inpetrol). Colourless crystalline solid (5.62 g, 0.011 mol, 83 %). ¹H NMR:(500 MHz, CDCl₃) δ d. 4.33 (9 H, ArCH₂NHCO₂C(CH₃)₃), NH), s. 2.37 (9H,ArCH₃), s. 1.44 (27H, CO₂C(CH₃)₃). ¹³C NMR: (125 MHz, CDCl₃) δ 155.7(CO₂C(CH₃)₃), 136.9, 133.5 (Ar), 79.7 (CO₂C(CH₃)₃), 40.0ArCH₂NHCO₂C(CH₃)₃), 28.5 (CO₂C(CH₃)₃), 16.0 (ArCH₃).

Compound 4c

Prepared by an analogous route to Compound 4b using Compound 3b (0.919g, 1.69 mmol) Purified by MPLC (0➔40 % EtOAc in petroleum ether).Colourless crystalline solid (0.655 g, 1.15 mmol 68 %). ¹H NMR: (4500MHz, CDCl₃) δ d 4.38 (6 H, ³J_(HH) = 4.2 Hz ArCH₂NHCO₂C(CH₃)₃), s. 3.79(9H, OCH₃) 1.43 (27H, CO₂C(CH₃)₃).

Compound 4d

Prepared in an analogous fashion to Compound 4b using Compound 3d (2.155g, 0.005 mol) was dissolved in MeOH (30 mL). Solid NaOH (0.599 g, 0.015mol). Purified by MPLC (50 % EtOAc in DCM) and then recrystalised fromDCM/petrol. Colourless crystalline solid (0.88 g, 0.002 mmol 40 %). ¹HNMR: (400 MHz, CDCl₃) δ s. 7.08 (1H, ArH) d. 4.24 (6 H, ³J_(HH) = 6.6Hz, ArCH₂N(O)OC(CH₃)₃), s. 1.46 (9 H, C(CH₃)₃). ¹³C NMR: (125 MHz,CDCl₃) δ 155.7 (CO₂C(CH₃)₃), 136.8 (Ar), 125.2 (Ar), 78.9 (CO₂C(CH₃)₃),45.2 (ArCH₂NHCO₂C(CH₃)₃), 28.2 (CO₂C(CH₃)₃).

Compound 5a

Method A: A pre-dried 250 mL round-bottomed flask fitted with a nitrogeninlet and reflux condenser and mounted in a heating block was chargedwith triphosgene (1.78 g, 6 mmol). The flask was placed under vacuum(ca. 10⁻² mbar) for 10 min the re-filled with nitrogen. Anhydroustoluene (150 mL) was added to give a colourless, homogenous solution. Tothis, a solution of 1,3,5-triaminomethyl-2,4,6-triethylbenzene (0.5 g, 2mmol) that had been previously dried by one azeotropic distillation oftoluene (80 mL), in anhydrous toluene (20 mL) was added via syringe over10 mins. After the addition was complete the temperature was raised to125° C. and the reaction allowed to reflux. After 75 min. the flask wasallowed to cool to room temperature and the solvent removed in a rotaryevaporator. The residue was extracted with toluene (3 × 10 mL) andconcentrated to dryness to give a pale-yellow oil that crystallised onstanding. Pale yellow crystalline solid (350 mg, 1.07 mmol, 54 %).

Method B: A pre-dried 200 mL Schlenk flask was charged with a magneticstirrer and Compound 4a (1.266 g, 2.25 mmol) and then placed under anitrogen atmosphere. 2-chloropyridine (1.7 mL, 20.21 mmol) and anhydrousDCM (70 mL) were added via syringe to give a colourless, homogenoussolution. Triflic anhydride (1.5 mL, 10.11 mmol) was added dropwise atambient temperature over 2 mins with stirring (400 rpm). The reactionwas stirred for 30 mins before a small aliquot of the reaction mixture(ca. 50 mL) was withdrawn and analysed by TLC (SiO₂, 50 % Et₂O inpetrol), which revealed complete consumption of the starting material(R_(f) = 0.24) and conversion to 5a (R_(f) = 0.5). The solvent wasremoved on a rotary evaporator to give an off-white solid. The solidswere extracted with Et₂O (× 2 15 mL) and passed through an alumina plug(20 mm × 20 mm), eluting with a further 20 mL of Et₂O. The colourlessfiltrate was evaporated to dryness and the residue recrystalised fromhexane. Colourless crystalline solid (0.435 g, 1.33 mmol, 59 %). ¹H NMR:(400 MHz, Toluene-d₆) δ s. 3.93 (6H, ArCH₂NCO), m. 2.51-2.37 (6H,ArCH₂CH₃), m. 0.96-0.86 (9H, ArCH₂CH₃). ¹³C NMR: (100 MHz, Toluene-d₆)143.2, 132.6 (Ar), 124.0 (NCO), 40.4 (ArCH₂NCO), 22.8 (ArCH₂CH₃), 16.0(ArCH₂CH₃).

Compound 5b

Prepared by an analogous route to Compound 5a (Method B) using Compound4c (0.351 g, 0.616 mmol) Colourless crystalline solid (0.144 g, 0.432mmol 70 %). ¹H NMR: (400 MHz, Toluene-d₈) δ d. 3.90 (6 H, ArCH₂NCO), s.3.40 (9 H, OCH₃). ¹³C NMR: (125 MHz, Toluene-d₈) δ 159.7 (Ar), 121.4(NCO), 62.9 (ArOCH₃), 36.2 (ArCH₂NCO).

Compound 5c

Prepared by an analogous route to Compound 5a (Method B) using Compound4b (0.300 g, 0.985 mmol) Colourless crystalline solid (0.198 g, 0.694mmol, 71 %). ¹H NMR: (500 MHz, Toluene-d₈) δ s. 3.73 (6 H, ArCH₂NCO), s.1.91 (9 H, CH₃). ¹³C NMR: (125 MHz, Toluene-d₈) δ 135.9, 133.4 (Ar),124.4 (ArCH₂NCO), 41.2 (ArCH₂NCO), 15.1 (ArCH₃).

Compound 5d

Prepared by an analogous route to Compound 5a (Method B) using Compound4d (0.250 g, 0.537 mmol). Colourless oil (0.062 g, 0.255 mmol, 48%). ¹HNMR: (500 MHz, Toluene-d₈) δ s. 6.56 (3 H, ArH), s. 3.64 (6 H,ArCH₂NCO). ¹³C NMR: (125 MHz, Toluene-d₈) δ 138.4 (Ar), 124.2 (Ar,ArCH₂NCO), 45.7 (ArCH₂NCO).

Compound 5e

A pre-dried round bottom flask was charged with1,3,5-triaminomethyl-2,4,6-triethylbenzene tristrifluoroacetate (0.535g, 0.905 mmol). THF (9 mL) was added, followed by CS₂ (1.100 mL, 18.100mmol) and DCC (0.585 mg, 2.806 mmol). The reaction was stirred for 16 hand then concentrated under vacuum. The resultant residue was trituratedwith DCM and the filtrate purified by MPLC (5% EtOAc/DCM → 40%) to givea white solid (0.168 g, 0.447 mmol, 49%). ¹H NMR: (400 MHz, CDCl₃) δ s.4.74 (6H, ArCH₂NCS), q. 2.84 (6H, J= 7.6 Hz, ArCH₂CH₃), t. 1.26 (9H, J =7.6 Hz, ArCH₂CH₃). ¹³C NMR: (100 MHz, CDCl₃) 144.2, 132.4 (Ar), 130.1(NCS), 42.9 (ArCH₂NCS), 23.2 (ArCH₂CH₃), 15.8 (ArCH₂CH₃).

Compound 5f

A pre-dried round bottom flask was charged with tris(2-aminotheyl)amine(5.000 g, 0.033 mol). THF (500 mL) was added, followed by CS₂ (40.0 mL,0.660 mol) and DCC (21.258 g, 0.102 mol). The reaction was stirred for16 h and then filtered. The filtrate was then concentrated under vacuum.The resultant residue was triturated with DCM and the filtrate purifiedby MPLC (12% EtOAc/DCM → 30%) to give a yellow solid (3.340 g, 0.023mmol, 71%). ¹H NMR: (500 MHz, CDCl₃) δ t. 4.74 (6H, J = 6.2 Hz,NCH₂CH₂NCS), t. 2.96 (6H, J = 6.2 Hz, NCH₂CH₂NCS). ¹³C NMR: (125 MHz,CDCl₃) 132.8 (NCS), 54.5 (NCH₂CH₂NCS), 44.4 (NCH₂CH₂NCS).

Compound 6b

A pre-dried Schlenk tube was charged with compound 5b (18 mg, 0.053mmol) and compound 84 (333 mg, 0.186 mmol) under a flow of nitrogen. DryTHF (3 mL) and anhydrous pyridine (0.024 mL, 0.053 mmol) was added. Aflake of MoO₂Cl₂ was added under a flow of nitrogen. The reaction wasstirred for 16 h. The reaction mixture was concentrated to dryness andthe crude residue purified by MPLC (2:3 EtOAc:DCM) to give a brown solid(174 mg, 0.030 mmol, 57%). ¹H NMR: (500 MHz, CDCl₃) δ m. 7.77-7.07 (33H,ArH), m. 4.56-4.06 (15H, ArCH_(2j), CO₂CH₂CH),), s. 3.66 (9H, ArOCH₃),m. 2.31-1.80z (144H, NHCH₂CH₂C(O)), s. 1.34 (162H, CO₂C(CH₃)₃), s. 1.33(81H, CO₂C(CH₃)₃). ¹³C NMR: (125 MHz, CDCl₃) δ 173.1, 173.0 (CONH),172.7, 172.7 (CO₂C(CH₃)₃), 172.6 (ArCONHR), 166.6 (NHC(O)NH), 158.4(COCH₃), 153.9 (CO₂CH₂CH), 143.8 (CNHFmoc), 143.6 (Fmoc 4°), 141.3(CNHFmoc), 141.2 (Fmoc 4°), 127.7 (Fmoc Ar), 127.7, 127.7 (CCO₂NHR),127.1 (Fmoc Ar), 127.0 (CNHC(O)NH), 125.3, 125.3 (CHCHCNH), 124.5 (FmocAr), 120.0 (Fmoc Ar), 119.9 (CHCNHFmoc), 119.9 (CCH₂NHC(O)NH), 80.5,80.4 (CO₂C(CH₃)₃), 67.3 (CO₂CH₂CH), 60.4 (ArOCH₃), 57.4, 57.4(C(CH₂CH₂CO₂)₃), 57.4 (C(CH₂CH₂CONH)₃), 47.2 (CO₂CH₂CH), 39.2(ArCH₂NHC(O)NH), 32.2 (CH₂CH₂CO₂C(CH₃)₃), 31.7 (CH₂CH₂CONH), 29.8(CH₂CH₂CONH), 29.7 (CH₂CH₂CO₂C(CH₃)₃), 28.0 (CO₂C(CH₃)₃).

Compound 6c

A pre-dried Schlenk tube was charged with compound 84 (441 mg, 0.245mmol) under a flow of nitrogen and a solution of compound 5c (20 mg,0.070 mmol) in THF (0.5 mL) added. Dry THF (3.5 mL) and anhydrouspyridine (0.006 mL, 0.070 mmol) was added. The reaction was stirred for16 h at 50° C. The reaction mixture was concentrated to dryness and thecrude residue purified by reverse phase MPLC on a 120 g SNAP Ultra C18cartridge elution (70-95% acetone/H₂O) to give a white solid (197 mg,0.035 mmol, 50%). ¹H NMR: (400 MHz, methanol-d₄) δ m. 7.86-7.03 (33H,ArH), m. 4.48-4.04 (15H, ArCH₂, CO₂CH₂CH),), s. 2.37 (9H, ArCH₃), m.2.31-1.86z (144H, NHCH₂CH₂C(O)), s. 1.40 (243H, CO₂C(CH₃)₃). ¹³C NMR:(125 MHz, methanol-d₄) δ 175.6, 175.5 (CONH), 174.4 (CO₂C(CH₃)₃), 174.4(ArCONHR), 157.1 (NHC(O)NH), 153.0 (CO₂CH₂CH), 145.0 (CNHFmoc), 142.6(Fmoc 4°), 134.4 (CCH₃), 131.2 (Fmoc Ar), 128.9 (CCO₂NHR), 128.2(CNHC(O)NH), 126.3 (CHCHCNH), 121.4 (Fmoc Ar), 121.1 (CHCNHFmoc), 121.1(CCH₂NHC(O)NH), 81.6 (CO₂C(CH₃)₃), 59.4, 58.8 (C(CH₂CH₂CO₂)₃), 58.7(C(CH₂CH₂CONH)₃), 40.1 (ArCH₂NHC(O)NH), 30.7 (CH₂CH₂CO₂C(CH₃)₃), 30.5(CH₂CH₂₋CONH), 28.5 (CH₂CH₂CONH, CH₂CH₂CO₂C(CH₃)₃), 28.4 (CO₂C(CH₃)₃),16.2 (ArCH₃).

Compound 6d

A pre-dried Schlenk tube was charged with compound 84 (441 mg, 0.245mmol) under a flow of nitrogen and a solution of compound 5d (212 mg,0.049 mmol) in THF (0.5 mL) was added. Dry THF (2.5 mL) and anhydrouspyridine (0.004 mL, 0.049 mmol) was added. The reaction was stirred for32 h. The reaction mixture was concentrated to dryness and the cruderesidue purified by reverse phase MPLC on a 120 g SNAP Ultra C18cartridge elution (70-95% acetone/H₂O) to give a white solid (110 mg,0.020 mmol, 40%). ¹H NMR: (500 MHz, methanol-d₄) δ m. 8.03-7.06 (33H,ArH), m. 4.55-4.17 (15H, ArCH₂, CO₂CH₂CH), m. 2.27-1.85 (144H,NHCH₂CH₂C(O)), s. 1.41 (243H, CO₂C(CH₃)₃). ¹³C NMR: (125 MHz,methanol-d₄) δ 175.5 (CONH), 174.4 (CO₂C(CH₃)₃), 174.4 (ArCONHR), 157.3(NHC(O)NH), 153.9 (CO₂CH₂CH), 143.8 (CNHFMoc), 143.6 (Fmoc 4°), 141.1(Fmoc 4°), 129.0 (CCO₂NHR), 127.7 (Fmoc Ar), 126.3 (CHCHCNH), 124.4(Fmoc Ar), 122.0 (CHCCH₂NHC(O)NH), 120.9 (CHCNHFmoc), 120.0 (Fmoc Ar),81.6 (CO₂C(CH₃)₃), 58.7 (C(CH₂CH₂CO₂)₃, C(CH₂CH₂CONH)₃), 40.7(CO₂CH₂CH), 40.1 (ArCH₂NHC(O)NH), 30.7 (CH₂CH₂CO₂C(CH₃)₃), 30.5(CH₂CH₂CONH), 28.5 (CH₂CH₂CONH, CH₂CH₂CO₂C(CH₃)₃), 28.5 (CO₂C(CH₃)₃).

Compound 6h-1 and 6h-2 Compound 6h-1

Compound 6h-2

A Schlenk flasked was charged with a stirrer bar, compound 84 (0.933 g,0.519 mmol) and compound 5a (0.100 g, 0.305 mmol) were dissolved inanhydrous THF (6 mL), pyridine (0.147 mL, 1.833 mmol) was added and themixture was then heated to 50° C. for 5 hours. Compound 11 (0.228 g,0.397 mmol) in anhydrous THF (1 mL) was added in one portion and thereaction stirred for a further 12 hours. The reaction mixture wastransferred to a RBF washing the Schlenk with CH₂Cl₂ beforeconcentrating under vacuum. The crude residue obtained was then purifiedby reverse phase flash chromatography on a 120 g SNAP Ultra C18cartridge elution (1CV 85% acetone/H₂O, 10 CV 85-95% acetone/H₂O, 2CV95% acetone) gave resolved peaks (fr17-29) 6h-1 (511 mg, 37%) and(fr8-15) 6h-2 (458 mg, 46%).

Compound 6h-1

¹H NMR: (400 MHz, (CD₃OD): δ 8.02-7.46 (19 H, br. m, ArH), 7.46-7.11 (14H, br. m, ArH), 4.60-4.30 (12 H, br. m, NHCH₂Ph and FmocH), 4.19 (3 H,br. s, NHCH₂Ph and FmocH), 3.71-3.55 (14 H, m, PEG CH₂), 3.33 (2 H, m,PEG CH₂), 2.85 (6 H, br. s, CH₂), 2.35-1.86 (96 H, m, dendrimer CH₂),1.42 (162 H, s, CH₃), 1.23 (9 H, br. s, CH₃); HRMS: (ESI⁺) calculatedfor C₂₄₄H₃₅₂N₂₁O₅₇Na₃ ²⁺: 1520.8407, found [M+3Na]³⁺: 1520.8395.

Compound 6h-2

¹H NMR: (400 MHz, (CD₃OD): δ 8.01-7.51 (19 H, br. m, ArH), 7.43-7.17 (14H, br. m, ArH), 4.54-4.25 (12 H, br. m, NHCH₂Ph and FmocH), 4.15 (3 H,br. s, NHCH₂Ph and FmocH), 3.69-3.47 (28 H, m, PEG CH₂), 3.26 (4 H, m,PEG CH₂), 2.82 (6 H, br. s, CH₂), 2.33-1.80 (48 H, m, dendrimer CH₂),1.42 (81 H, s, CH₃), 1.17 (9 H, br. s, CH₃);

Compound 6i

Into a dry Schlenk equipped with a stirrer bar, compound 5a (55.0 mg,0.168 mmol) and compound 2 (480 mg, 0.588 mmol) were weighed. AnhydrousTHF (3.3 mL) was added. The reaction mixture was stirred at 50 degreesfor 20 hours, then transferred to a round bottom flask and concentratedto dryness. The crude product was purified by MPLC, on a 60 g C18 SNAPULTRA cartridge, elution (3CV 70% acetone/ water, 10CV 70- 95% acetone/water, 3CV 95% acetone) produced a resolved peak (fr18-22). White solid273 mg, 59%. ¹H NMR: (400 MHz, CD₃OD) δ 7.05- 7.85 (33H, m, ArH), 4.35(3H, s, Flu-CH), 4.26 (6H, s, Flu-CHCH₂O), 3.01 (18H, m, H16), 2.71 (6H,m, ArCH₂CH₃), 1.93 (18H, m, H15), 1.30 (81H, s, ^(t)Bu), 1.06 (9H, t, J=7.00 Hz, ArCH₂CH₃). ¹³C NMR: (101 MHz, CD₃OD) δ 15.59 (9C, ArCH₂GH₃),22.23(3C, ArCH₂CH₃), 27.31 (27C, ^(t)Bu), 34.89 (9C, CCH₂CH₂NH), 35.21(9C, CCH₂CH₂NH), peak not observed (1C, Flu- CHCH2), 51.32 (1C,C(CH₂CH₂NHBoc)₃), peak not observed (9C, ^(t)Bu), peak not observed (1C,Flu- CH), 119.60- 127.47 (60C, Ar), 141.28 (9C, Boc C=O), 143.64 (1C,Fmoc C=O), 156.89 (1C, Ar-C=O).

Compound 7b

To a stirred solution of 6b (0.170 g, 0.030 mmol) was in DCM (5 mL) at0° C., was added distilled DBU (0.006 mL, 0.040 mmol). The reaction wasstirred at 0° C. for 2 h before concentrating under vacuum. The cruderesidue obtained was then purified by reverse phase flash chromatographyon a 120 g SNAP Ultra C18 cartridge elution (1CV 80% acetone/H₂O, 10 CV80-95% acetone/H₂O, 2CV 95% acetone) to give an off-white solid (0.133g, 0.026 mmol, 89%). ¹H NMR: (500 MHz, methanol-d₄) δ m. 7.30-7.27 (3H,CHCNH (Ar)), m. 7.22-7.15 (6H, CHCHCNH, CHCNH₂ (Ar)), s. 4.54 (6H,ArCH₂NH), s. 3.89 (9H, ArOCH₃), m. 2.25-1.91 (144H, NHCH₂CH₂C(O)), s.1.43 (243H, CO₂C(CH₃)₃). ¹³C NMR: (125 MHz, methanol-d₄) δ 175.5, 175.4(CONH), 174.4, 174.3 (CO₂C(CH₃)₃), 170.6, 170.0 (ArCONHR), 160.2(NHC(O)NH), 157.9 (COCH₃), 141.4, 140.7 (CNH₂), 134.7, 132.7 (CCO₂NHR),129.9, 125.6 (CNHC(O)NH), 124.4, 123.6, 120.7, 118.8 (CHCHCNH), 117.2,116.6 (CHCNH₂), 115.5 (CCH₂NHC(O)NH), 81.6 (CO₂C(CH₃)₃), 59.3 (ArOCH₃),58.7, 58.6 (C(CH₂CH₂CO₂)₃), 54.8 (C(CH₂CH₂CONH)₃), 35.0, 35.0(ArCH₂NHC(O)NH), 32.7, 32.5 (CH₂CH₂CO₂C(CH₃)₃), 32.2, 32.2 (CH₂CH₂CONH),30.7 (CH₂CH₂CONH), 30.4, (CH₂CH₂CO₂C(CH₃)₃), 28.5, 28.4 (CO₂C(CH₃)₃).

Compound 7c

Prepared in a manner analogous to 7b from compound 6c (0.097 g, 0.017mmol). Purified by reverse phase flash chromatography on a 120 g SNAPUltra C18 cartridge elution (1CV 80% acetone/H₂O, 10 CV 80-95%acetone/H₂O, 2CV 95% acetone) to give a white solid (0.072 g, 0.014mmol, 85%). ¹H NMR: (500 MHz, methanol-d₄) δ m. 7.42-7.30 (3H, CHCNH(Ar)) m. 7.31-7.29 CHCNH₂(Ar)), m. 7.23-7.19 (3H, CHCHCNH (Ar)), m.4.54-4.48 (6H, ArCH\₂NH), s. 2.49 (6H, ArCH₃), s. (3H, ArCH₃), m.2.28-1.90 (144H, NHCH₂CH₂C(O)), s. 1.44 (243H, CO₂C(CH₃)₃). ¹³C NMR:(125 MHz, methanol-d₄) δ 175.5 (CONH), 174.4, 174.3 (CO₂C(CH₃)₃), 170.1(ArCONHR), 158.1 (NHC(O)NH), 141.3 (CNH₂), 135.4 (CCO₂NHR), 134.7(CNHC(O)NH), 130.0 (CCH₃), 124.4, 118.9 (CHCHCNH), 117.4 (CHCNH₂), 111.4(CCH₂NHC(O)NH), (C(CH₂CH₂CONH)₃), 81.6 (CO₂C(CH₃)₃),58.7 (G(CH₂CH₂CO₂)₃,C(CH₂CH₂CONH)₃), 40.3 (ArCH₂NHC(O)NH), 32.5 (CH₂CH₂CO₂C(CH₃)₃), 32.2(CH₂CH₂CONH), 30.7 (CH₂CH₂CONH), 30.5, (CH₂CH₂CO₂C(CH₃)₃), 28.4(CO₂C(CH₃)₃),16.3 (ArCH₃).

Compound 7d

Prepared in a manner analogous to 7b from compound 6d (0.110 g, 0.020mmol). Purified by reverse phase flash chromatography on a 120 g SNAPUltra C18 cartridge elution (1CV 80% acetone/H₂O, 10 CV 80-95%acetone/H₂O, 2CV 95% acetone) to give a white solid (0.022 g, 0.004mmol, 21%). ¹H NMR: (500 MHz, methanol-d₄) δ m. 7.43-7.13 (12H, Ar), m.4.50-4.29 (6H, ArCH₂NH), m. 2.34-1.86 (144H, NHCH₂CH₂C(O)), s. 1.43(243H, CO₂C(CH₃)₃). ¹³C NMR: (HSQC) (125 MHz, methanol-d₄) δ 175.5(CONH), 174.4 (CO₂C(CH₃)₃),174.4 (ArCONHR), 157.3 (NHC(O)NH), 141.2(CNH₂), 135.4 (CCO₂NHR), 134.7 (CNHC(O)NH), 126.3 (CHCHCNH), 124.4,(CHCHCNH), 122.0 (CHCCH₂NHC(O)NH), 118.9 (CHCHCNH), 115.6 (CHCNH₂), 81.6(CO₂C(CH₃)₃),58.7 (C(CH₂CH₂CO₂)₃, C(CH₂CH₂CONH)₃), 44.4 (ArCH₂NHC(O)NH),35.1 (CH₂CH₂CO₂C(CH₃)₃), 34.6 (CH₂CH₂CONH), 30.4 (CH₂CH₂CONH), 30.2,(CH₂CH₂CO₂C(CH₃)₃), 28.0 (CO₂C(CH₃)₃).

Compound 7e

Compound 13 (231 mg, 0.30 mmol) was dried by azeotrope with toluene,then under an inert N₂ atmosphere was dissolved in anhydrousdichloromethane (1.5 mL). Pyridine (41 uL, 0.51 mmol) was added,followed by a solution of compound 5a (8 mg, 0.086 mmol) in anhydrousdichloromethane (0.5 mL) and the reaction mixture was stirred at 34° C.until complete by TLC. The solvent was removed under vacuum and thecrude product purified by flash column chromatography (SiO₂,EtOAC:CH₂Cl₂ 1:4 to 1:2 then MeOH:CH₂Cl₂ 5:95) to afford theFMOC-protected intermediate. HRMS: (ESI+) calculated forC₁₅₀H₁₉₂N₁₂Na₃O₃₀ ³⁺ 983.7859, found [M+3Na]³⁺: 983.7844. Under an inertN₂ atmosphere, the FMOC-protected intermediate (140 mg, 0.053 mmol) wasdissolved in anhydrous dichloromethane (9 mL) and cooled to 0° C. DBU(50 µL, 0.34 mmol) was added dropwise and the reaction mixture warmed toroom temperature and stirred for 1 hour. The solvent was removed undervacuum and the crude product purified by flash column chromatography(SiO₂, CH₂Cl₂ then 7.5% MeOH:CH₂Cl₂) to afford compound 7e (95 mg, 0.048mmol, 91%). ¹H NMR: (400 MHz, CD₃OD, 298 K): δ 7.40 (s, 3H, NH), 7.35(d, J = 8.2 Hz, 3H, H₁₁), 7.21 (d, J = 2.0 Hz, 3H, H₈), 7.10 (dd, J =8.2, 2.0 Hz, 3H, H₁₀), 4.52 (s, 6H, H₅), 2.94 - 2.87 (m, 6H, H₂), 2.30 -2.25 (m, 18H, H₁₆), 2.12 - 2.06 (m, 18H, H₁₅), 1.45 (s, 81H, H₁₉), 1.26(t, J = 7.4 Hz, 9H, H₁); HRMS: (ESI+) calculated for C₁₀₅H₁₆₄N₁₂O₂₄ ²⁺989.1002, found [M+2H]²⁺ _(:) 989.1004.

Compound 11

Prepared in an analogous fashion to compound 2 by treating compound A1(3.308 g, 5.443 mmol), (3.308 g, Moles: 5.443 mmol) was added anddissolved in S1 (Vol: 22.680 mL). R3 (Vol: 1.270 mL, Moles: 7.258 mmol)was added dropwise followed by R2 (Vol: 1.000 mL, Moles: 4.536 mmol) andthe reaction was left to stir until complete by TLC. TLC 70%EtOAc/CH₂Cl₂ showed the reaction to be complete at 24 h (visualised byUV/weak ninhydrin stain). The reaction mixture was transferred to aRBF - washing with CH₂Cl₂ and concentrated under vacuum to yield a brownresidue. The crude product was purified by flash column chromatographyeluting with 50% to 100% EtOAc/CH₂Cl₂. Colourless amorphous solid (2.50g, 4.35 mmol, 80 %). ¹H NMR: (400 MHz, CDCl₃) δ 7.74 (d, J = 7.5 Hz, 2H,ArH), 7.60 (d, J= 2.0 Hz, 2H, ArH), 7.45 (s, 1H, ArH), 7.38 (t, J= 7.5Hz, 2H, ArH), 7.26 (d, J= 9.6 Hz, 2H, ArH), 7.05 (s, 1H, ArH), 6.84 (s,1H ArH), 6.65 (d, J= 8.4 Hz, 1H C(O)NHCH₂), 4.49 (s, 1H, Flu-CH₂), 4.18(s, 2H, Flu-OCH₂), 3.68 - 3.45 (m, 14H, OCH₂), 3.27 (t, J = 5.0 Hz, 2HC(O)NHCH₂), 1.34 (d, J = 6.7 Hz, 2H, N₃CH₂).

Compound 12

Prepared in an analogous fashion to compound 2 by treating compound A1(2.000 g, 3.291 mmol) in 16.5 mL DCM and DIPEA (0.917 mL, 5.266 mmol)with propargyl amine (0.422 mL, Moles: 6.583 mmol) for 72 hours.Purified by column chromatography using ethyl acetate as the eluent.White solid (1.36 g, 3.290 mmol, 99 %). ¹H NMR: (400 MHz, Methanol-d₄) δ7.95 (d, J = 7.5 Hz, 2H, ArH), 7.89 - 7.80 (m, 3H, ArH), 7.61 (d, J= 8.4Hz, 1H, ArH), 7.52 (t, J= 7.4 Hz, 2H, ArH), 7.44 (s, 2H), 6.87 (d, J=8.4 Hz, 1H, ArH), 4.45 (s, 2H), 4.40 (s, 1H, Flu-CH₂), 4.17 (d, J= 2.5Hz, 2H, Flu-OCH₂), 2.87 (t, J= 2.4 Hz, 1H, CH₂CCH), 2.67 (p, J= 1.9 Hz,2H, CH₂CCH).

Compound 13

A 50 mL RBF was charged with compound A1 (500 mg, 1.017 mmol),di-tert-butyl-4-amino-4-(3-(tert-butoxy)-3-oxopropyl)heptanedioate* (560mg, 1.35 mmol) (560 mg, 1.35 mmol) and anhydrous toluene (10 mL). Theslurry was evaporated to dryness and the residue re-dissolved inanhydrous pyridine (5 mL) and DCM (3 mL). The mixture was stirred at 50°C. for 16 hours. The solvent was removed to give a viscous brown oil,which was partitioned between EtOAc and 1 M aq HCl. The organic phasewas washed with water then brine. The combined organic fractions wereconcentrated and then absorbed onto silica gel and purified by flashchromatograohy (EtOAc:DCM (20→50 %). Pink powder (467 mg, 0.612 mmol, 60%). ¹H NMR: (400 MHz, CDCl₃) δ 7.79 (d, J= 7.6 Hz, 2H, ArH), 7.64 (d, J=2.1 Hz, 1H, ArH), 7.43 (t, J = 7.5 Hz, 2H, ArH), 7.34 (s, 3H, ArH), 6.78(d, J= 9.0 Hz, 1H, NH), 6.60 (s, 1H, ArH), 6.30 (s, 1H, ArH), 4.56 (s,2H, Flu-CH₂O), 4.28 (s, 1H, Flu-CH₂), 4.08 (s, 2H), 2.30 (dd, J = 8.8,6.7 Hz, 6H, CH₂C(O)), 2.16 - 2.04 (m, 6H, CCH₂), 1.44 (s, 24H, C(CH₃)₃).

* Newkome, George R.; Weis, Claus D. Organic Preparations and ProceduresInternational, 1996, vol. 28, #4 p. 495 - 498

Compound 14

A Schlenk flash was charged with (3S,4S)-pyrrolidine-3,4-diol (100.0 mg,0.970 mmol), di-tert-butyl4-(3-(tert-butoxy)-3-oxopropyl)-4-isocyanatoheptanedioate (385.4 mg,0.873 mmol) and anhydrous DMF under N₂ to give orange solution. Thesolution was left to stir for 16 hours the poured into water (10 mL) andextracted with EtOAc (10 mL). The organic layer was separated, dried andconcentrated to dryness. Off white solid. (430 mg, 0.79 mmol, 81 %). ¹HNMR: (400 MHz, CDCl₃) δ 4.65 (s, 1H, C(O)NH), 4.15 - 4.06 (m, 2H, CHOH),3.95 (s, 2H, OH), 3.52 (dd, J= 10.9, 4.2 Hz, 2H, NCH₂), 3.21 (d, J= 10.8Hz, 2H, NCH₂), 2.17 (dd, J= 9.0, 6.7 Hz, 6H, CH₂C(O)), 2.02 - 1.82 (m,6H, CCH₂), 1.37 (s, 27H, C(CH₃)₃). ¹³C NMR: (125 MHz, CDCl₃) δ 176.8 (s,NC(O)N), 168.1 (s, CCO₂C), 79.7 (s, CO₂C(CH₃)₃), 75.7 (s, COH), 51.5 (s,CH₂N), 30.8 (s, CH₂C(O)), 29.8 (s, CCH₂), 28.1 (s, CO₂C(CH₃)₃).

Compound 15

A Schlenk flash was charged with Compound 14 (400.0 mg, 0.734 mmol)anhydrous DCM and TEA (0.409 mL, 2.93 mmol) and neat methylsulfonylchloride (0.125 mL, 1.62 mmol) was added dropwise. The orange solutionwas allowed to stir overnight to give turbid orange solution. TLC (SiO₂,100% EtOAc) indictaed complete consumption of the starting material to anew product (Rf = 0.4). Water was added (10 mL) and the organic layerseparated, dried with MgSO₄ and concentrated to dryness. Orangecrystalline solid (428 mg, 0.611 mmol, 83 %). ¹H NMR: (400 MHz, CDCl₃) δ5.42 (s, 1H, C(O)NH), 5.2 (d, J = 4.5, 2H, CHOS(O₂)O), 3.77 (dd, J=12.2, 4.5 Hz, 2H, NCH₂), 3.65 (d, J= 12.2 Hz, 2H, NCH₂), 3.11, (s, 1H,CH₃S(O)2O), 2.24 (t, J = 8.0 Hz, 6H, CH₂C(O)), 1.97 (m, 6H, CCH₂), 1.42(s, 27H, C(CH₃)₃). ¹³C NMR: (125 MHz, CDCl₃) δ 176.9 (s, NC(O)N), 168.3(s, CCO₂C), 79.8 (s, CO₂C(CH₃)₃), 79.0 (s, CH₃S(O)₂OC), 49.0 (CH₂N),38.6 (s, CH₃S(O)20), 30.5 (s, CH₂C(O)), 30.2 (s, CCH₂), 28.4 (s,CO₂C(CH₃)₃).

Compound 15a

Compound 15 (428.0 mg, 0.611 mmol), dissolved in dry DMF and solid NaN₃(119.2 mg, 1.833 mmol, 3.000 eq) added in one portion. The orangesuspension was heated to 100° C. for 16 h to give a dark brown solution.Partitioned between EtOAc and water, washed with x2 10 mL 5 % LiCl(aq),brine (1 × 10 mL), dried over MgSO₄ then concentrated to dryness withca. 1 g silica gel. This silica absorbed crude product was loaded onto afrit on top of 1 cm x 2 cm plug of fresh silica and eluted with 50 %EtOAc in petrol (ca. 20 mL). The colourless filtrate was evaporated todryness. Colouress crystalline solid (256 mg, 0.43 mmol, 74 %). ¹H NMR:(400 MHz, CDCl₃) δ 5.21 (s, 1H, C(O)NH), 3.92 (m, 2H, CHN₃), 3.60 (dd, J= 10.9, 5.7 Hz, 2H, NCH₂), 3.30 (ds, J = 10.9, 3.3 Hz, 2H, NCH₂), 2.19(t, J = 7.5 Hz, 6H, CH₂C(O)), 1.92 (m, 6H, CCH₂), 1.37 (s, 27H,C(CH₃)₃). ¹³C NMR: (125 MHz, CDCl₃) δ 176.8 (s, NC(O)N), 169.3 (s,CCO₂C), 79.8 (s, CO₂C(CH₃)₃), 64.0 (s, CN₃), 48.5 (s, CH₂N), 30.3 (s,CH₂C(O)), 29.9 (s, CCH₂), 28.3 (s, CO₂C(CH₃)₃).

Compound 16

Compound 15a (200.0 mg, 0.336 mmol) was dissolved in EtOH (1.000 mL) andmixed with 10 % Pd/C (25.0 mg, 0.235 mmol). The reaction flask waspurged nitrogen and then a hydrogen. The resulting mixture was stirredfor 20 hours under a hydrogen atmosphere (supplied from a balloon). Thecrude reaction mixture was filtered through Celite™ washing andconcentrated under reduced pressure. The crude product was purified byflash column chromatography (SiO₂, MeCN:CH₂Cl₂ 1:4 then MeOH:CH₂Cl₂ 1:9)to afford compound 16 (93 mg, 0.171 mmol, 51%). ¹H NMR: (400 MHz, CDCl₃,298 K): δ 4.59 (s, 1H, NH), 3.63 (dd, J = 10.1, 5.3 Hz, 2H, H₃), 3.08(q, J = 5.7, 5.0 Hz, 2H, H₉), 2.94 (dd, J = 10.3, 5.7 Hz, 2H, H₃), 2.17(t, J = 7.8 Hz, 6H, H₄), 1.95 - 1.85 (m, 6H, H₅), 1.38 (s, 27H, H₁). ¹³CNMR: (100 MHz, CDCl₃, 298 K): δ 173.18 (C₃), 155.63 (C₇), 80.57 (C₂),58.37 (C₉), 56.74 (C₆), 52.03 (C₈), 30.53 (C₅), 29.91 (C₄), 28.13 (C₁);HRMS: (ESI⁺) calculated for C₂₇H₅₀N₄NaO₇ ⁺ 565.3572, found [M+Na]⁺:565.3547.

Compound 200 - 1,3,5-tris(bromomethyl)-2-bromo-4,6-dimethylbenzene

2-bromo-4,6-dimethylbenzene (5.000 g, 0.027 mol, 1.000 eq),paraformaldehyde (12.736 g, 0.424 mol, 15.700 eq) and AcOH/HBr 33% (70mL) were added to a dry 200 mL round bottom flask. The mixture wasstirred while ZnBr₂ (15.211 g, 0.068 mol, 2.500 eq) was slowly added andthe mixture was heated to 90° C. After 24 hours, an additional portionparaformaldehyde (12.736 g, 0.424 mol, 15.700 eq) and 2.7 g ZnBr₂(12.736 g, 0.424 mol, 15.700 eq) was added. The yellow solution washeated an additional. 140 hours. The reaction mixture was then cooled toroom temperature and the colourless crystals isolated by filtration,washing with AcOH (3 x 10 mL) and then water until the pH of thefiltrate was neutral. Dried under vacuum for 2 day to afford colourlesscrystals (7.50 g, 0.016 mol, 60 %).

¹H NMR (400 MHz, CDCl₃) δ 4.78 (s, 4H, CH₂), 4.54 (s, 2H, CH₂), 2.54 (s,6H, CH₃).

Compound 201 - 1,3,5-tris(azidomethyl)-2-bromo-4,6-dimethylbenzene

1,3,5-tris(bromomethyl)-2-bromo-4,6-dimethylbenzene (1.326 g, 2.859mmol) was dissolved in dry DMF (30.000 mL) under nitrogen and stirredwhile NaN₃ (1.115 g, 17.154 mmol). The reaction mixture was heated to40° C. and stirred overnight. The reaction was cooled and poured intowater (100 mL) and ppt extracted with EtOAc (3x50 mL). The combinedorganic fractions were washed with 5 % LiCl (2 x 20 mL) and then brine,dried (MgSO₄) and concentrated to dryness behind a blast shield toafford a colourless oil the crystallised on standing. (1.00 g, 2.856mmol, 99 %).

¹H NMR (400 MHz, CDCl₃) δ 4.72 (s, 4H, CH₂), 4.55 (s, 2H, CH₂), 2.50 (s,6H, CH₃).

Compound 202 - 1,3,5-tris(aminomethyl)-2-bromo-4,6-dimethylbenzeneTrihydrochloride

1,3,5-tris(azidomethyl)-2-bromo-4,6-dimethylbenzene (0.600 g, 1.713mmol, 1.000 eq) and triphenylphopshine (3.011 g, 11.48 mmol, 6.7) weredissolved in THF (12 mL) and water (0.375 mL, 20.78 mmol, 12.13 eq)added. Stirred at 40° C. overnight. Solvent removed completely andtreated with 0.5 M HCl. (10 mL). Extracted with EtOAc (2 x 10 mL) andthe water layered reserved and concentrated to near dryness the added torapidly stirred acetone (20 mL). The white precipitate was collected ona frit and washed with acetone (10 mL) and the dried under vacuum.Colourless solid (0.65 g, 1.71 mmol, 99 %).

¹H NMR (400 MHz, D₂O) δ 4.54 (s, 4H, CH₂), 4.38 (s, 2H, CH₂), 2.53 (s,6H, CH₃).

Compound 203 - Di-tert-butyl((2-bromo-5-(((tert-butoxycarbonyl)amino)methyl)-4,6-dimethyl-1,3-phenylene)bis(methylene))dicarbamate

1,3,5-tris(aminomethyl)-2-bromo-4,6-dimethylbenzene trihydrochloride(0.68 g, 1.78 mmol), BOC₂O (2.33 g, 10.7 mmol) and triethylamine (1.5mL, 10.7 mmol) was dissolved in MeOH (70 mL) and stirred at roomtemperature overnight. The solvent was removed and the residue portionedbetween EtOAc (50 mL) and 0.5 M citric acid. The aqueous layer wasextracted with EtOAc (2x20 mL) and the combined organic fractionscombined, dried (MgSO₄) and concentred to dryness to yield a colourlessoil which crystallised upon standing (0.80 g, 1.4 mmol, 78 %)

¹H NMR (400 MHz, CDCl₃) δ 4.77 (br. s, 2H, NH), 4.55 (d, J = 5.7 Hz, 4H,CH₂), 4.36 (br. s, 3H, NH and CH₂), 2.48 (s, 6H, CH₃), 1.44 (s, 9H,CH₃);

Compound 204 - 1-bromo-2,4,6-tris(isocyanatomethyl)-3,5-dimethylbenzene

Prepared by an analogous route to Compound 103 using Compound 202 (0.20g, 0.35 mmol) to give Compound 204 as a light pink crystalline solid (94mg, 0.27 mmol, 77%).

-   ¹H NMR (400 MHz, CDCl₃) δ 4.70 (s, 4H, CH₂), 4.49 (s, 2H, CH₂), 2.52    (s, 6H, CH₃);-   ¹³C NMR (101 MHz, CDCl₃) δ 138.4 (C), 135.1 (C), 134.4 (C), 129.1    (C), 124.4 (C), 45.6 (CH₂), 41.3 (CH₂), 16.7 (CH₃);-   HRMS (ESI⁺) calculated for C₁₄H₁₃BrN₃O₃ ⁺: requires 350.0135, found    [M+H]⁺: 350.0139.

Compound 205

A Schlenk was dried under vacuum by heat-gun and allowed to cool to RT.Compound 84 (628.4 mg, 0.350 mmol, 3.5 eq) was added to the flask anddissolved in anhydrous toluene (5 mL). The solvent was removed on theline to azeotropically dry the reagent and the resulting residue left todry for 30 mins prior to use. The solid was re-dissolved in anhydroustoluene (4 mL) and a solution of1-bromo-2,4,6-tris(isocyanatomethyl)-3,5-dimethylbenzene (Compound 204,35.0 mg, 0.100 mmol, 1.0 eq) in anhydrous toluene (1 mL) was added tothe flask. Pyridine (0.048 mL, 0.600 mmol, 6.0 eq) was added and thereaction heated to 34° C. for 24 h. (N.B. the reaction became a vibrantorange colour overtime). TLC (5% MeOH/CH₂Cl₂, UV and Seebachvisualisation) indicated that the reaction was complete. The reactionwas concentrated under vacuum and purified by reverse phase flashchromatography (on a SNAP Ultra C18 120 g cartridge, 70:30 Acetone:Waterto 100:0 over 12 CV). Fraction 6-11 contained excess 3-G2MM Linker whichwas recovered and fraction 15-18 contained Compound 205 as an off-whitefoam (348 mg, 61%).

¹H NMR (400 MHz, CD₃OD) δ 8.09 - 7.49 (br. m, 19H, ArH), 7.51 - 7.10(br. m, 14H, ArH), 4.74 - 4.54 (br. m, 4H, benzylic CH₂ and FmocH),4.51 - 4.05 (br. s, 11H, benzylic CH₂ and FmocH), 2.50 (br. s, 6H,methyl CH₃), 2.31 - 1.81 (m, 144H, dendrimer CH₂), 1.41 (s, 243H,dendrimer CH₃);

HRMS (Nanospray ESI⁺) calculated for C₃₀₈H₄₆₆BrN₂₁O₇₅ ⁴⁺: requires1435.8134, found [M+4H]⁴⁺ _(:) 1435.8147.

Compound 206

Compound 205 (327.0 mg, 0.057 mmol, 1.000 eq) was dissolved in anhydrousCH₂Cl₂ (2.9 mL) and cooled to 0° C. DBU (0.051 mL, 0.342 mmol, 6.0 eq)was added dropwise and the reaction stirred for 2 h. The reactionmixture was then concentrated under vacuum and purified by reverse phaseflash chromatography (on a SNAP Ultra C18 60 g cartridge, 70:30Acetone:Water to 100:0 over 12 CV) to give the Compound 206 as a lightpink solid (0.278 g, 97%).

-   ¹H NMR (400 MHz, CD₃OD) δ 7.40 (dd, J= 8.3, 3.5 Hz, 3H, ArH), 7.30    (d, J= 2.1 Hz, 3H, ArH), 7.21 (dd, J = 8.3, 2.1 Hz, 3H, ArH), 4.73    (br. s, 4H, benzylic CH₂), 4.52 (br. s, 2H, benzylic CH₂), 2.60 (br.    s, 6H, CH₃), 2.31 - 2.14 (m, 72H, dendrimer CH₂), 2.13 - 2.04 (m,    18H, dendrimer CH₂), 2.00 - 1.88 (m, 54H, dendrimer CH₂), 1.44 (s,    243H, dendrimer CH₃);-   ¹³C NMR (126 MHz, CD₃OD) δ 175.5 (C), 174.4 (C), 170.1 (C), 158.1    (C), 158.0 (C), 141.4 (C), 140.2 (C), 137.1 (C), 136.1 (C), 132.8    (C), 130.7 (C), 130.0 (C), 129.9 (C), 124.4 (CH), 118.9 (CH), 117.3    (CH), 81.7 (C), 59.4 (C), 58.7 (C), 32.5 (CH₂), 32.2 (CH₂), 30.7    (CH₂), 30.5 (CH₂), 28.5 (CH₃), 17.1 (CH₃);-   HRMS (Nanospray ESI⁺) calculated for C₂₆₃H₄₃₅BrN₂₁O₆₉ ³⁺. requires    1691.0137, found [M+3H]³⁺ _(:) 1691.0132.

Compound 207

Dissolved Compound 206 (270.0 mg, 0.053 mmol, 1.0 eq) in anhydrouspyridine (22.3 mL) and heated the reaction mixture to 40° C. in a drysyn, external temperature of heat probe set to 40° C. In a separatepear-shaped flask TEB NCO (Compound 103, 20.9 mg, 0.064 mmol, 1.2 eq)was dissolved in anhydrous CH₂Cl₂ (2.3 mL). The TEB NCO solution wasadded by syringe pump at 0.85 mL/h. Once the addition was complete thereaction was left overnight at 40° C. The reaction mixture wasconcentrated under vacuum on a liquid nitrogen cold finger rotaryevaporator to dryness. The resulting foam was co-evaporated with toluenetwice and the resulting foam purified by reverse phase chromatography(loading in MeCN and eluting on a SNAP Ultra C18 60 g cartridge startingat Acetone:Water 70:30 to 100:0 over 12 CV). Fraction 3-6 containedCompound 207 as a colourless foam (196 mg, 68%).

-   ¹H NMR (500 MHz, CD₃OD) δ 8.20 - 7.83 (m, 4H, ArH), 7.73 - 7.56 (m,    5H, ArH), 7.44 (s, 5H, NH), 4.55 - 4.31 (m, 12H, benzylic CH₂),    2.96 - 2.74 (m, 6H, ethyl CH₂), 2.55 (s, 6H, methyl CH₃), 2.36 -    1.80 (m, 144H, dendrimer CH₂), 1.44 (s, 243H, dendrimer CH₃), 1.25 -    1.16 (m, 9H, ethyl CH₃);-   ¹³C NMR (126 MHz, CD₃OD) δ 175.5, 175.4, 174.3, 169.5, 169.3, 158.3,    158.0, 157.9, 157.5, 144.4, 144.4, 139.7, 136.9, 136.2, 135.2,    134.3, 134.1, 132.7, 132.0, 131.4, 130.7, 129.8, 125.9, 125.8,    125.1, 124.5, 122.6, 81.6, 59.5, 59.4, 58.8, 58.7, 43.5, 40.1, 38.9,    32.4, 32.2, 32.2, 30.7, 30.7, 30.4, 28.5, 23.6, 17.1, 16.9, 16.7;-   HRMS (Nanospray ESI⁺) calculated for C₂₈₁H₄₅₅BrN₂₄O₇₂Na³⁺: requires    1808.0615, found [M+2H+Na]³⁺: 1808.0601.

Compound 208 - Receptor 4

Dissolved Compound 207 (196 mg, 0.036 mmol, 1.0 eq) in CH₂Cl₂ (9.0 mL)and TFA (2.4 mL) was added. The reaction was left overnight at RT andadded dropwise to 300 mL of H₂O to precipitate the acid. This suspensionwas centrifuged in 50 mL batches and then washed and sonicated with H₂O.The isolated solid was then dried under high vacuum to give Compound 208as a colourless solid (108 mg, 77%).

¹H NMR (500 MHz, DMSO-d₆) δ 12.04 (s, 27H, COOH), 8.23 - 7.66 (m, 6H,ArH), 7.59 - 7.41 (m, 3H, ArH), 7.41 - 7.16 (m, 6H, NH), 6.67 - 6.34 (m,6H, NH), 4.45 - 4.20 (m, 12H, benzylic CH₂), 2.81 (s, 6H, ethyl CH₂),2.47 (s, 6H, methyl CH₃), 2.28 - 1.75 (m, 144H, dendrimer CH₂), 1.20 -1.08 (m, 9H, ethyl CH₃);

¹³C NMR (126 MHz, DMSO-d₆) δ 174.4, 172.4, 165.7, 158.5, 158.3, 156.0,155.6, 154.8, 150.6, 142.9, 138.0, 135.4, 135.1, 134.7, 133.9, 133.5,133.1, 130.2, 129.7, 129.1, 122.9, 78.7, 57.4, 56.4, 30.8, 30.4, 29.0,28.1 22.4, 16.7, 16.2.

Compound 209

Prepared in an analogous fashion to Compound 2 by treating HBTUactivated linker (Compound A1, 14.3 g, 11.8 mmol, 1.0 eq) in CH₂Cl₂(16.5 mL) and DIPEA (2.7 mL, 21 mmol, 1.8 eq) with propargyl amine (1.6mL, 25 mmol, 2.1 eq) for 72 hours. Purified by column chromatographyusing ethyl acetate as the eluent to give Compound 209 as colourlesssolid (2.24 g, 5.4 mmol, 46%).

¹H NMR (400 MHz, CD₃OD) δ 7.87 - 7.60 (m, 5H, ArH), 7.57 - 7.45 (m, 1H,ArH), 7.45 - 7.23 (m, 4H, ArH), 6.79 (d, J = 8.4 Hz, 1H, ArH), 4.45 (br.s, 2H, Fmoc CH₂), 4.27 (br. s, 1H, Fmoc CH), 4.11 (d, J= 2.5 Hz, 2H,alkyne CH₂), 2.56 (p, J= 2.5 Hz, 1H, alkyne CH).

HRMS (ESI⁺) calculated for C₂₅H₂₁N₃O₃Na⁺: requires 434.1475, found[M+Na]⁺: 434.1479.

Compounds 210a and 210b Compound 210a

Compound 210b

A Schlenk flasked was charged with a stirrer bar, Compound 84 (0.933 g,0.519 mmol, 1.7 eq) and Compound 5a (0.100 g, 0.305 mmol, 1.0 eq) anddissolved in anhydrous THF (6 mL) and pyridine (0.145 mL, 1.833 mmol,6.0 eq) then heated to 50° C. for 5 h. Compound 209 (0.189 g, 0.458mmol, 1.5 eq) was added in one portion and the reaction stirred for afurther 12 h. The reaction mixture was transferred to an RBF washing theSchlenk with CH₂Cl₂ before concentrating under vacuum. The crude residueobtained was then purified by reverse phase flash chromatography (loadedwith MeCN) on a 120 g SNAP Ultra C18 cartridge elution (1CV 85%acetone/H₂O, 10 CV 85-95% acetone/H₂O, 2CV 95% acetone) to give(fr18-24) identified as Compound 210b a colourless solid (312 mg, 35%),(fr37-47) identified as Compound 210a a colourless solid (524 mg, 40%)and (fr57-60) identified as Compound 108 as colourless solid (354 mg,20%).

¹H NMR Compound 210a (400 MHz, CDCl₃) δ 8.06 — 7.54 (m, 19H, ArH), 7.48— 7.11 (14H, m, ArH), 4.60 — 4.28 (m, 13H, benzylic CH₂ and FmocH), 4.23— 4.09 (m, 4H, benzylic CH₂, FmocH and alkyne CH₂), 2.84 (br. s, 6H,ethyl CH₂), 2.61 (t, J = 2.5 Hz, 1H, alkyne CH), 2.31 — 1.87 (m, 96H,dendrimer CH₂), 1.43 (s, 162H, dendrimer CH₃), 1.20 (br. s, 9H, ethylCH₃);

HRMS Compound 210a (Nanospray ESI⁺) calculated for C2₃₉H₃₄₂N₁₈O₅₄Na₂ ²⁺:requires 2188.2210, found [M+2Na]²⁺: 2188.2224.

¹H NMR Compound 210b (400 MHz, CDCl₃) δ 8.01 — 7.51 (m, 19H, ArH), 7.47— 7.07 (m, 14H, ArH), 4.61 — 4.27 (m, 13H, benzylic CH₂ and FmocH), 4.21— 4.08 (m, 4H, benzylic CH₂, FmocH and alkyne CH₂), 2.84 (br. s, 6H,ethyl CH₂), 2.61 (t, J = 2.5 Hz, 2H, alkyne CH), 2.34 — 1.82 (m, 48H,dendrimer CH₂), 1.44 (s, 81H, dendrimer CH₃), 1.19 (br. s, 9H, ethylCH₃);

HRMS Compound 210b (Nanospray ESI⁺) calculated for C₁₆₆H₂₁₃N₁₅O₃₃Na₂ ²⁺:requires 1495.7634, found [M+2Na]²⁺: 1495.7628.

Compound 211

A RBF charged with Compound 210a (0.524 g, 0.121 mmol, 1.0 eq) dissolvedin anhydrous CH₂Cl₂ (6 mL) under nitrogen was cooled to 0° C. DBU (0.108mL, 0.726 mmol, 6.0 eq) was added dropwise and the reaction mixture wasleft for 2 h at 0° C. The reaction mixture was then concentrated undervacuum and purified by normal phase flash chromatography on a SNAPKP-Sil 50 g cartridge (eluting with CH₂Cl₂:MeOH 100:0 to 90:10 over 12CV). This material was then purified by reverse phase chromatography(loaded with MeCN) on a SNAP Ultra C18 60 g cartridge (eluting withAcetone:Water 75:25 to 100:0 over 12 CV) to give Compound 211 as acolourless solid (0.200 g, 0.055 mmol, 45%).

¹H NMR (500 MHz, CD₃OD) δ 7.69 — 7.54 (m, 1H, ArH) 7.46 — 7.32 (m, 3H,ArH), 7.33 — 7.26 (m, 2H, ArH), 7.23 — 7.19 (m, 2H, ArH), 7.17 — 7.12(m, 1H, ArH), 4.54 — 4.43 (m, 6H, benzylic CH₂) 4.11 (d, J = 2.5, 2H,alkyne CH₂), 2.88 (m, 6H, ethyl CH₂), 2.58 (t, J = 2.5, 1H, alkyne CH),2.32 — 2.03 (m, 60H, dendrimer CH₂), 2.01 — 1.84 (m, 36H, dendrimerCH₂), 1.43 (s, 162H, dendrimer CH₃), 1.25 (m, 9H, ethyl CH₃);

HRMS (Nanospray ESI⁺) calculated for C₁₉₄H₃₁₅N₁₈O₄₈ ³⁺: requires1222.4271, found [M+3H]³⁺: 1222.4281.

Compound 212

A RBF charged with Compound 210b (0.312 g, 0.106 mmol, 1.0 eq) dissolvedin anhydrous CH₂Cl₂ (5.3 mL) under nitrogen was cooled to 0° C. DBU(0.095 mL, 0.635 mmol, 6.0 eq) was added dropwise and the reactionmixture was left for 2 h at 0° C. The reaction mixture was thenconcentrated under vacuum and purified by normal phase flashchromatography on a SNAP KP-Sil 50 g cartridge eluting (CH₂Cl₂:MeOH100:0 to 90:10 over 12 CV). This material was then purified by reversephase chromatography (loaded with MeCN) on a SNAP Ultra C18 60 gcartridge (eluting with Acetone:Water 75:25 to 100:0 over 12 CV) to giveCompound 212 as a colourless solid (0.130 g, 0.057 mmol, 54%).

¹H NMR (500 MHz, CD₃OD) δ 7.71 — 7.50 (m, 1H, ArH) 7.46 - 7.32 (m, 3H,ArH), 7.31 - 7.22 (m, 2H, ArH), 7.22 - 7.16 (m, 1H, ArH), 7.16 - 7.06(m, 2H, ArH), 4.61 - 4.34 (m, 6H, benzylic CH₂) 4.11 (br. s, 4H, alkyneCH₂), 2.88 (q, J = 7.5, 6H, ethyl CH₂), 2.58 (t, J = 2.5, 2H, alkyneCH), 2.33 - 2.04 (m, 30H, dendrimer CH₂), 2.02 - 1.86 (m, 18H, dendrimerCH₂), 1.44 (s, 81H, dendrimer CH₃), 1.24 (t, J = 7.5, 9H, ethyl CH₃);

HRMS (Nanospray ESI⁺) calculated for C₁₂₁H₁₈₅N₁₅O₂₇ ²⁺: requires1140.6798, found [M+2H]²⁺: 1140.6805.

Compound 213

Dissolved Compound 211 (200.0 mg, 0.055 mmol, 1.0 eq) in anhydrouspyridine (22.9 mL) and heated the reaction to 40° C. in a dry syn,external temperature of heat probe set to 40° C. In a separatepear-shaped flask Compound 5a (21.6 mg, 0.066 mmol, 1.2 eq) wasdissolved in anhydrous CH₂Cl₂ (2.3 mL). The Compound 5a solution wasadded by syringe pump at 0.85 mL/h. Once the addition was complete thereaction was left overnight at 40° C. The reaction mixture wasconcentrated under vacuum on a liquid nitrogen cold finger rotaryevaporator to dryness. The resulting foam was co-evaporated with toluenetwice and the resulting foam purified by reverse phase chromatography(loading in MeCN and eluting on a SNAP Ultra C18 60 g cartridge startingat Acetone:Water 70:30 to 100:0 over 12 CV). Fraction 5-9 containedCompound 213 as a colourless foam (115 mg, 53%).

-   ¹H NMR (500 MHz, CD₃OD) δ 8.07 - 7.94 (m, 6H, ArH), 7.68 (d, J = 8.4    Hz, 2H, ArH), 7.63 (d, J = 8.4 Hz, 1H, ArH), 7.46 (s, 1H, NH),    4.59 - 4.34 (m, 12H, benzylic CH₂), 4.17 (d, J = 2.5 Hz, 2H, alkyne    CH₂), 3.00 - 2.85 (m, 6H, ethyl CH₂), 2.85 - 2.74 (m, 6H, ethyl    CH₂), 2.63 (t, J = 2.5 Hz, 1H, alkyne CH), 2.36 - 2.09 (m, 60H,    dendrimer CH₂), 1.97 (t, J = 8.2 Hz, 36H, dendrimer CH₂), 1.45 (s,    162H, dendrimer CH₃), 1.26 - 1.18 (m, 18H, dendrimer CH₃);-   ¹³C NMR (126 MHz, CD₃OD) δ 175.5, 175.5, 174.4, 169.3, 169.0, 158.4,    158.3, 157.2, 157.2, 144.5, 144.5, 144.4, 137.1, 136.9, 134.3,    134.2, 134.0, 133.8, 131.2, 130.1, 130.1, 129.4, 126.1, 125.9,    125.5, 125.3, 122.7, 122.3, 81.6, 80.8, 72.2, 59.4, 58.8, 58.7,    38.9, 38.8, 38.8, 38.7, 32.4, 32.2, 30.7, 30.4, 28.4, 23.6, 16.8,    16.7, 16.6;-   HRMS (Nanospray ESI⁺) calculated for C₂₁₂H₃₃₆N₂₁O₅₁ ³⁺: requires    1331.4802, found [M+3H]³⁺: 1331.4784.

Compound 214 - Receptor 5

Dissolved Compound 213 (111 mg, 0.028 mmol, 1.0 eq) in CH₂Cl₂ (7.0 mL)and TFA (1.9 mL) was added. The reaction was left overnight at RT andadded dropwise to 300 mL of H₂O to precipitate the acid. This suspensionwas centrifuged in 50 mL batches and then washed and sonicated with H₂O.The isolated solid was then dried under high vacuum to give Compound 214as a colourless solid (36 mg, 43%).

¹H NMR (500 MHz, DMSO-d₆) δ 12.02 (s, 27H, COOH), 8.73 (s, 1H, NH),8.19 - 7.67 (m, 6H, ArH), 7.64 - 7.12 (m, 9H, ArH and NH), 6.48 (s, 3H,NH), 6.39 (s, 3H, NH), 4.52 - 4.15 (m, 12H, benzylic CH₂), 4.11 - 3.93(m, 2H, alkyne CH₂), 3.34 (s, 6H, NH), 3.09 (s, 1H, alkyne CH), 2.82 (s,6H, ethyl CH₂), 2.66 (s, 6H, ethyl CH₂), 2.28 - 1.62 (m, 144H, dendrimerCH₂), 1.26 - 1.01 (m, 18H, ethyl CH₃);

¹³C NMR (126 MHz, DMSO-d₆) δ 174.9, 172.9, 166.0, 165.9, 156.4, 156.1,155.0, 142.8, 142.7, 135.5, 135.4, 133.9, 133.8, 133.4, 133.3, 129.2,128.7, 128.4, 127.5, 124.6, 123.9, 123.6, 123.1, 119.9, 119.3, 82.1,73.1, 57.9, 56.8, 37.6, 37.2, 31.2, 30.8, 29.5, 28.9, 28.6, 22.8, 22.5,16.8, 16.7.

Compound 215

Dissolved Compound 212 (130.0 mg, 0.057 mmol, 1.0 eq) in anhydrouspyridine (23.8 mL) and heated the reaction to 40° C. in a dry syn,external temperature of heat probe set to 40° C. In a separatepear-shaped flask. Compound 5a (22.0 mg, 0.068 mmol, 1.2 eq) wasdissolved in anhydrous CH₂Cl₂ (2.4 mL). The Compound 5a solution wasadded by syringe pump at 0.85 mL/h. Once the addition was complete thereaction was left overnight at 40° C. The reaction mixture wasconcentrated under vacuum on a liquid nitrogen cold finger rotaryevaporator to dryness. The resulting foam was co-evaporated with toluenetwice and the resulting foam purified by reverse phase chromatography(loading in MeCN and eluting on a SNAP Ultra C18 60 g cartridge startingat Acetone:Water 70:30 to 100:0 over 12 CV). Fraction 3-4 containedCompound 215 as a colourless foam (33 mg, 23%).

-   ¹H NMR (500 MHz, CD₃OD) δ 8.03 (d, J = 2.1 Hz, 2H, ArH), 7.99 - 7.92    (m, 4H, ArH), 7.66 (dd, J = 8.6, 2.1 Hz, 1H, ArH), 7.61 (dd, J =    8.6, 2.1 Hz, 2H, ArH), 7.45 (s, 1H, NH), 4.56 -4.36 (m, 12H,    benzylic CH₂), 4.15 (d, J = 2.5 Hz, 4H, alkyne CH₂), 2.97 - 2.84 (m,    6H, ethyl CH₂), 2.84 - 2.72 (m, 6H, ethyl CH₂), 2.61 (t, J= 2.5 Hz,    2H, alkyne CH), 2.32 - 2.09 (m, 30H, dendrimer CH₂), 2.01 - 1.88 (m,    18H, dendrimer CH₂), 1.43 (s, 81H, dendrimer CH₃), 1.28 - 1.13 (m,    18H, ethyl CH₃);-   ¹³C NMR (126 MHz, CD₃OD) δ 175.6, 175.5, 174.4, 169.4, 169.1, 158.4,    158.3, 157.3, 157.2, 144.5, 144.5, 144.5, 137.2, 136.9, 134.3,    134.2, 133.9, 133.8, 131.1, 130.1, 130.1, 129.3, 126.1, 126.0,    125.5, 125.3, 122.7, 122.2, 81.7, 80.8, 72.1, 59.5, 58.8, 58.7,    38.9, 38.8, 38.7, 38.7, 32.4, 32.2, 32.2, 30.7, 30.4, 30.0, 28.4,    23.6, 16.7, 16.6, 16.5;-   HRMS (Nanospray ESI⁺) calculated for C₁₃₉H₂₀₆N₁₈O₃₀ ²⁺: requires    1304.2589, found [M+2H]²⁺: 1304.2606.

Compound 216 - Receptor 6

Dissolved Compound 215 (30 mg, 0.012 mmol, 1.0 eq) in CH₂Cl₂ (3.0 mL)and TFA (0.81 mL) was added. The reaction was left overnight at RT andadded dropwise to 300 mL of H₂O to precipitate the acid. This suspensionwas centrifuged in 50 mL batches and then washed and sonicated with H₂O.The isolated solid was then dried under high vacuum to give Compound 216as a colourless solid (17 mg, 67%).

¹H NMR (500 MHz, DMSO-d₆) δ 12.03 (s, 27H, COOH), 8.75 (t, J = 5.7 Hz,1H, NH), 8.21 - 7.93 (m, 6H, ArH), 7.91 - 7.73 (m, 4H, NH), 7.60 - 7.42(m, 3H, ArH), 7.38 - 7.21 (m, 6H, ArH and NH), 6.46 (s, 3H, NH), 6.37(s, 3H, NH), 4.52 - 4.21 (m, 12H, benzylic CH₂), 4.03 (d, J = 2.5 Hz,2H, alkyne CH₂), 3.10 (t, J = 2.5 Hz, 1H, alkyne CH), 2.83 (s, 6H, ethylCH₂), 2.69 (m, 6H, ethyl CH₂), 2.18 - 1.72 (m, 144H, dendrimer CH₂),1.19 - 1.09 (m, 18H, ethyl CH₂);

¹³C NMR (126 MHz, DMSO-d₆) δ 174.9, 172.9, 166.0, 165.9, 156.3, 156.1,155.0, 149.3, 142.8, 142.8, 137.6, 135.5, 135.4, 133.9, 133.8, 133.3,130.6, 129.2, 128.7, 128.3, 127.5, 124.7, 124.6, 123.9, 123.7, 123.1,120.4, 119.9, 119.3, 82.1, 73.1, 57.8, 56.8, 37.5, 37.3, 31.2, 30.8,29.5, 28.9, 28.5, 22.8, 22.5, 16.8, 16.7.

Compound 217

N—Boc—L—serine (5.00 g, 24.4 mmol, 1.0 eq) was dissolved in anhydrousDMF (50 mL), cooled to 0° C. and NaH (2.05 g, 51.1 mmol, 2.1 eq) (60%dispersion in mineral oil) was added. After stirring for 30 min at 0°C., 3-bromopropyne (2.52 mL, 26.7 mmol) (80% solution in toluene) wasadded dropwise. After stirring at 0° C. for 30 minutes the ice bath wasremoved and stirring continued overnight at ambient temperature. Thesolution was dark brown in colour after this time. Aqueous sulfatebuffer (16 g Na₂SO₄, 2 mL H₂SO₄ made up to 150 mL with H₂O) was added tothe flask gradually to avoid any exotherm generated upon quenching. Aprecipitate formed and then slowly dissolved to give an orange solution.Brine (150 mL) was added and the reaction mixture extracted with EtOAc(3 × 150 mL). The combined organics were washed with H2O (3 × 150 mL),dried over MgSO4, filtered and the resulting filtrate concentrated undervacuum. Some DMF was transferred over during this step. Before theorganic had completely concentrated silica gel was added and thenconcentrated to dryness. The silica was then washed with CH₂Cl₂ (500 mL)before eluting the desired compound with 10% MeCN/CH₂Cl₂ (500 mL) oruntil fractions did not contain the product (PMA stain, 10% MeCN/CH₂Cl₂,streaky black spot 0.3-0.4 R_(f)). The resulting filtrate wasconcentrated under vacuum to give the desired compound as a yellow gumafter drying under high vacuum (4.9 g, 83%).(S)-2-[(tert-butoxycarbonyl)amino]-3-(prop-2-yn-1-yloxy)propanoic acid(2.1 g, 8.6 mmol, 1.0 eq) was dissolved in CH₂Cl₂ (33.6 mL) and TFA(33.1 mL) was added. The reaction was stirred at RT until complete byTLC. The reaction mixture was concentrated under vacuum and azeotropewith toluene to give the TFA salt of O-(prop-2-yn-1-yl)serine which wastaken directly onto the next step. Dissolved O-(prop-2-yn-1-yl)serine(2.22 g, 8.63 mmol, 1.0 eq) in acetone (14.4 mL) and H₂O (14.4 mL).Sodium carbonate (2.75 g, 25.9 mmol, 3.0 eq) and Fmoc—OSu (3.06 g, 9.07mmol, 1.05 eq) was added and the reaction left to stir overnight. Thereaction mixture was acidified to pH 3 with HCl (3 M) and extracted withEtOAc (3 × 100 mL). The combined organic phases were dried over Na₂SO₄and the resulting filtrate concentrated under vacuum. The crude residuewas purified by column chromatography (eluting with 2.5% MeOH in CH₂Cl₂)to give N-(((9H-fluoren-9-yl)methoxy)carbonyl)-O-(prop-2-yn-1-yl)serineas a white solid (2.83 g, 90%). Second generation dendritic amine(Compound 82, 600 mg, 0.417 mmol, 1.0 eq) was dissolved in THF (3.6 mL).N-(((9H-fluoren-9-yl)methoxy)carbonyl)-O-(prop-2-yn-1-yl)serine (183 mg,0.500 mmol, 1.2 eq), COMU (0.21 g, 0.50 mmol, 1.2 eq), K-Oxyma (90 mg,0.50 mmol, 1.2 eq) and DIPEA (0.22 mL, 1.25 mmol, 3.0 eq) were thenadded to the solution. The reaction was left to stir overnight at RT.The reaction was concentrated under vacuum to remove THF. The residuewas redissolved in EtOAc (40 mL) and then washed sequentially with KHSO₄(100 mL), sat. NaHCO₃ (100 mL) and brine (100 mL). The organic phase wasdried over MgSO₄, filtered and concentrated under vacuum to give a crudeyellow oil. Purification by reverse phase flash chromatography on a SNAPUltra C18 120 g cartridge (eluting with Acetone:Water 78:22 for 3CV then91:9 for 4CV) gave the Fmoc-protected intermediate (fr10-13) as an offwhite solid (602 mg, 81%). To a RBF dried under vacuum by heat-gun wasadded Fmoc-protected intermediate (1.2 g, 0.67 mmol, 1.0 eq). AnhydrousCH₂Cl₂ (33.6 mL) was added and the flask cooled to 0° C. DBU (0.20 mL,1.3 mmol, 2.0 eq) was added dropwise and the reaction stirred untilcomplete by TLC (5% MeOH/CH₂Cl₂, stain with ninhydrin to visualise). Thereaction flask was warmed to RT after 2 h. The reaction was concentratedunder vacuum and the crude residue purified by reverse phasechromatography on a SNAP Ultra C18 120 g cartridge (eluting withAcetone:Water 75:25 to 100:0 over 12 CV). Fr5-18 contained Compound 217as a yellow foam (0.95 g, 91%).

¹H NMR (400 MHz, CD₃OD) δ 4.58 (br. s, 1H, NH), 4.30 - 4.15 (m, 2H,alkyne CH₂), 3.72 (dd, J = 9.2, 5.4 Hz, 1H, serine CHH), 3.63 (dd, J =9.2, 5.4 Hz, 1H, serine CHH), 3.48 (t, J = 5.4 Hz, 1H, serine CH), 2.90(t, J = 2.4 Hz, 1H, alkyne CH), 2.29 - 1.90 (m, 48H, dendrimer CH₂),1.45 (s, 81H, dendrimer CH₃);

HRMS (ESI⁺) calculated for C₈₂H₁₄₂N₅O₂₃Na²⁺: requires 793.9991, found[M+H+Na]²⁺: 793.9974.

Compound 218

A RBF was dried under vacuum using a heat-gun. Once cool, Compound 217(0.45 g, 0.29 mmol, 1.0 eq) and HBTU activated linker (0.21 g, 0.35mmol, 1.2 eq) were added and suspended in anhydrous THF (1.44 mL). DIPEA(0.08 mL, 0.46 mmol, 1.6 eq) was then added and the reaction stirredovernight at RT. The reaction was heterogenous initially and became ahomogeneous (dark brown) solution after 16 h at RT. The reaction wasconcentrated under vacuum and purified by reverse phase flashchromatography on a SNAP Ultra C18 120 g cartridge (eluting withAcetone:Water 66:34 for 3CV then 88:12 for 5CV). Fr7-11 contained theCompound 218 as a yellow solid (441 mg, 80%).

¹H NMR (400 MHz, CD₃OD) δ 7.80 - 7.42 (m, 6H, ArH), 7.41 - 7.07 (m, 4H,ArH), 6.72 (d, J = 8.5 Hz, 1H, ArH), 4.47 (d, J = 5.3 Hz, 1H, serineCH), 4.39 (br. s, 2H, Fmoc CH₂), 4.19 (br. s, 1H, Fmoc CH), 4.17 - 4.07(t, J = 2.4 Hz, 2H, alkyne CH₂), 3.83 (dd, J = 9.6, 5.3 Hz, 1H, serineCHH), 3.75 (dd, J = 9.6, 5.3 Hz, 1H, serine CHH), 2.81 (t, J = 2.4 Hz,1H, alkyne CH), 2.17 - 1.73 (m, 48H, dendrimer CH₂), 1.32 (s, 81H,dendrimer CH₃);

HRMS (ESI⁺) calculated for C₁₀₄H₁₅₇N₇O₂₆Na₂ ²⁺: requires 983.5497, found[M+2Na]²⁺: 983.5496.

Compound 219

Compound 218 (0.64 mg, 0.33 mmol, 3.5 eq) was concentrated into apear-shaped flask and dried through azeotropic distillation withanhydrous toluene (5 mL) followed by drying under high vacuum for 30min. The resulting residue was dissolved in anhydrous CH₂Cl₂ (4.7 mL),Compound 5a (31 mg, 0.095 mmol, 1.0 eq) and pyridine (0.046 mL, 0.57mmol, 6.0 eq) were added and the reaction heated to 34° C. for 12 h. Thereaction mixture was concentrated under vacuum and the resulting residuepurified by flash column chromatography on a SNAP Ultra C18 120 gcartridge (eluting with Acetone:Water 70:30 to 95:5 over 12CV). Fr1-8recovered Compound 218 and fr11-14 contained Compound 219 as anoff-white solid (476 mg, 83%).

¹H NMR (400 MHz, CD₃OD) δ 7.93 - 7.36 (m, 19H, ArH), 7.34 - 7.05 (m,14H, ArH), 4.49 (t, J = 5.0 Hz, 3H, serine CH), 4.38 (br. s, 6H,benzylic CH₂), 4.27 (br. s, 6H, Fmoc CH₂), 4.15 (t, J = 2.2 Hz, 6H,alkyne CH₂), 4.07 (s, 3H, Fmoc CH), 3.84 (dd, J = 9.6, 5.0 Hz, 3H,serine CHH), 3.76 (dd, J = 9.6, 5.0 Hz, 3H, serine CHH), 2.81 (t, J =2.2 Hz, 1H, alkyne CH), 2.74 (br. s, 6H, ethyl CH₂), 2.22 - 1.71 (m,144H, dendrimer CH₂), 1.32 (s, 243H, dendrimer CH₃), 1.09 (t, J = 7.4Hz, 9H, ethyl CH₃).

HRMS (Nanospray ESI⁺) calculated for C₃₃₀H₄₉₆N₂₄O₈₁ ⁴⁺: requires1523.6382, found [M+4H]⁴⁺: 1523.6409.

Compound 220

A RBF charged with Compound 219 (0.451 g, 0.074 mmol, 1.0 eq) dissolvedin anhydrous CH₂Cl₂ (3.7 mL) under nitrogen and cooled to 0° C. DBU(0.066 mL, 0.444 mmol, 6.0 eq) was added dropwise and the reactionmixture was left for 2 h at 0° C. The reaction mixture was thenconcentrated under vacuum and purified by reverse phase flashchromatography on a SNAP Ultra C18 60 g cartridge (eluting withAcetone:Water 70:30 to 100:0 over 12 CV). Fr3-7 contained Compound 220as a colourless solid (0.279 g, 69%).

¹H NMR (400 MHz, CD₃OD) δ 7.37 (d, J = 8.3 Hz, 3H, ArH), 7.24 (d, J =2.1 Hz, 3H, ArH), 7.15 (dd, J = 8.2, 2.1 Hz, 3H, ArH), 4.48 (t, J = 5.3Hz, 3H, serine CH), 4.42 (br. s, 6H, benzylic CH₂), 4.25 - 4.10 (m, 6H,alkyne CH₂), 3.84 (dd, J = 9.6, 5.3 Hz, 3H, serine CHH), 3.76 (dd, J =9.6, 5.3 Hz, 3H, serine CHH), 2.83 (t, J = 2.4 Hz, 3H, alkyne CH), 2.80(q, J = 6.1 Hz, 6H, ethyl CH₂), 2.20 - 1.73 (m, 144H, dendrimer CH₂),1.34 (s, 243H, dendrimer CH₃), 1.22 -1.12 (m, 9H, ethyl CH₂).

¹³C NMR (126 MHz, CD₃OD) δ 175.4, 174.4, 171.6, 170.4, 157.8, 145.1,141.4, 133.9, 131.4, 130.5, 124.4, 119.0, 117.4, 81.7, 80.4, 77.0, 70.4,59.4, 59.4, 58.7, 56.2, 39.3, 32.2, 32.1, 30.7, 30.5, 28.5, 23.9, 17.0;

HRMS (Nanospray ESI⁺) calculated for C₂₈₅H₄₆₆N₂₄O₇₅ ⁴⁺: requires1357.0870, found [M+4H]⁴⁺: 1357.0824.

Compound 221

Compound 220 (265.0 mg, 0.049 mmol, 1.0 eq), DMAP (18 mg, 0.147 mmol,3.0 eq) and n-octyl glucoside (29 mg, 0.098 mmol, 2.0 eq) were weighedinto a round bottom flask and anhydrous toluene was added. This wasremoved in situ under high vacuum. Procedure repeated, and the resultingfoam allowed to dry for 30 min. The reagents were then dissolved inanhydrous CH₂Cl₂ (98 mL) and heated to 34° C. In a separate dried RBF,Compound 5a (19.2 mg, 0.059 mmol, 1.2 eq) was weighed into a flask anddissolved in anhydrous CH₂Cl₂ (9.8 mL). This solution was then syringepumped into the reaction mixture at 1 mL/hr. After completion, thereaction was left for a further 24 h at 34° C. The reaction mixture wascooled and concentrated under vacuum. Purification by reverse phaseflash chromatography on a SNAP Ultra C18 120 g column (eluting withAcetone:Water 70:30 to 100:0 over 12 CV). Fr1-8 were taken and purifiedby prep HPLC (C18 20x150 mm, 5 µm, 20 mL/min, Acetone:Water 70:30 to100:0 over 30 min). Analysis at this stage was difficult because of thepresence of n-octyl glucose (110 mg, 39%). Mass spec confirmed materialobtained after prep HPLC contained Compound 221.

HRMS (Nanospray ESI⁺) calculated for C₃₀₃H₄₈₇N₂₇O₇₈ ⁴⁺: requires1438.6254, found [M+4H]⁴+_(:) 1438.6232.

Compound 222 - Receptor 7

Compound 221 (110 mg, 0.019 mmol, 1.0 eq) was dissolved in CH₂Cl₂ (4.8mL) and TFA (1.3 mL) was added. The reaction was left overnight at RTand then concentrated under vacuum. Purification by reverse phase flashchromatography on a SNAP Ultra C18 30 g column (eluting withMeOH:Water+0.1 % formic acid 10:90 to 100:0 over 12 CV). Fr30-32 weretaken and purified by prep HPLC (C18 20x150 mm, 5 µm, 20 mL/min,MeOH:Water+0.1% formic acid 10:90 to 100:0 over 30 min) gave Compound222 as a colourless solid (30 mg, 37%).

¹H NMR (400 MHz, CD₃OD) δ 8.46 (s, 3H, NH), 8.17 (s, 3H, ArH), 8.02 (d,J = 8.5 Hz, 3H, ArH), 7.62 (d, J = 8.5 Hz, 3H, ArH), 4.65 (t, J = 5.4Hz, 3H, serine CH), 4.53 -4.33 (m, 12H, benzylic CH₂), 4.37 - 4.23 (m,6H, alkyne CH₂), 4.10 - 3.84 (m, 6H, serine CH₂), 2.98 (t, J = 2.4 Hz,3H, alkyne CH), 2.76 (s, 6H, ethyl CH₂), 2.69 (s, 6H, ethyl CH₂), 2.48 -1.68 (s, 144H, dendrimer CH₂), 1.41 - 1.00 (m, 18H, ethyl CH₃);

¹³C NMR (126 MHz, CD₃OD) δ 182.8, 175.2, 175.0, 170.6, 170.0, 160.8,157.4, 156.8, 144.2, 132.3, 131.9, 128.3, 127.8, 124.4, 68.7, 58.3,58.2, 54.7, 37.6, 31.6, 30.8, 30.6, 22.6, 22.3, 15.4.

Compound 223

A pre-dried Schlenk tube was charged with Compound 84 (441 mg, 0.245mmol) under a flow of nitrogen and a solution of Compound 5c (20 mg,0.070 mmol) in THF (0.5 mL) added. Dry THF (3.5 mL) and anhydrouspyridine (0.006 mL, 0.070 mmol) was added. The reaction was stirred for16 h at 50° C. The reaction mixture was concentrated to dryness and thecrude residue purified by reverse phase MPLC on a 120 g SNAP Ultra C18cartridge elution (70-95% acetone/H₂O) to give a Compound 223 whitesolid (197 mg, 0.035 mmol, 50%).

HRMS (Nanospray ESI⁺) calculated for C₃₀₉H₄₆₉N₂₁O₇₅ ⁴⁺: requires1419.3407, found _([M+4H]) ⁴+_(:) 1419.3391.

Compound 224

Prepared in a manner analogous to Compound 7b from Compound 223 (0.097g, 0.017 mmol). Purified by reverse phase flash chromatography on a 120g SNAP Ultra C18 cartridge elution (1CV 80% acetone/H₂O, 10 CV 80-95%acetone/H₂O, 2CV 95% acetone) to give a white solid (0.072 g, 0.014mmol, 85%).

¹H NMR (500 MHz, methanol-d₄) δ m. 7.42-7.30 (3H, CHCNH (Ar)) m.7.31-7.29 CHCNH₂ (Ar)), m. 7.23-7.19 (3H, CHCHCNH (Ar)), m. 4.54-4.48(6H, ArCH₂NH), s. 2.49 (6H, ArCH₃), s. (3H, ArCH₃), m. 2.28-1.90 (144H,NHCH₂CH₂C(O)), s. 1.44 (243H, CO₂C(CE₃)₃).

¹³C NMR (125 MHz, methanol-d₄) δ 175.5 (CONH), 174.4, 174.3(CO₂C(CH₃)₃), 170.1 (ArCONHR), 158.1 (NHC(O)NH), 141.3 (CNH₂), 135.4(CCO₂NHR), 134.7 (CNHC(O)NH), 130.0 (CCH₃), 124.4, 118.9 (CHCHCNH),117.4 (CHCNH₂), 111.4 (CCH₂NHC(O)NH), (C(CH₂CH₂CONH)₃), 81.6(CO2C(CH₃)₃), 58.7, (G(CH₂CH₂CO₂)₃), 40.3 (ArCH₂NHC(O)NH), 32.5(CH_(2G)H₂CO₂C(CH₃)₃), 32.2 (CH₂CH₂CONH), 30.7 (CH₂CH₂CONH), 30.5,(CH₂CH₂CO₂C(CH₃)₃), 28.4 (CO₂C(GH₃)₃), 16.3 (ArCH₃).

Compound 225

Dissolved Compound 224 (200 mg, 0.040 mmol, 1.0 eq) in anhydrouspyridine (16.8 mL) and heated the reaction to 40° C. in a dry syn,external temperature of heat probe set to 40° C. In a separatepear-shaped flask Compound 5c (13.7 mg, 0.048 mmol, 1.2 eq) wasdissolved in anhydrous CH₂Cl₂ (1.7 mL). The Compound 5c solution wasadded by syringe pump at 0.85 mL/h. Once the addition was complete thereaction was left overnight at 40° C. The reaction mixture wasconcentrated under vacuum on a liquid nitrogen cold finger rotaryevaporator to dryness. The resulting foam was co-evaporated with toluenetwice and the resulting foam purified by reverse phase chromatography(loading in MeCN and eluting on a SNAP Ultra C18 60 g cartridge startingat Acetone:Water 70:30 to 100:0 over 12 CV). Fraction 4-6 containedCompound 225 as a colourless foam (127 mg, 60%).

¹H NMR (500 MHz, CD₃OD) δ 8.07 (s, 2H, NH), 7.95 (s, 3H, ArH), 7.89 (br.s, 3H, ArH), 7.66 (d, J = 8.6 Hz, 3H, ArH), 7.44 (s, 6H, NH), 4.51 (s,6H, benzylic CH₂), 4.45 (s, 1H, benzylic CH₂), 2.49 (s, 9H, methyl CH₃),2.43 (s, 9H, methyl CH₃), 2.32 - 1.91 (m, 144H, dendrimer CH₂), 1.43 (s,243H, dendrimer CH₃).

¹³C NMR (126 MHz, CD₃OD) δ 175.5, 174.4, 169.4, 158.6, 157.6, 137.5,137.4, 135.1, 134.7, 131.4, 129.9, 126.0, 122.6, 81.6, 59.5, 58.8, 40.2,40.0, 32.5, 32.2, 30.7, 30.5, 28.5, 16.7, 16.3.

HRMS (Nanospray ESI⁺) calculated for C₂₇₉H₄₅₀N₂₄O₇₂Na₄ ⁴⁺: requires1345.7981, found [M+4Na]⁴⁺: 1345.7994.

Compound 226 — Receptor 8

Dissolved Compound 225 (120 mg, 0.023 mmol, 1.0 eq) in CH₂Cl₂ (5.8 mL)and TFA (1.5 mL) was added. The reaction was left overnight at RT andadded dropwise to 300 mL of H₂O to precipitate the acid. This suspensionwas centrifuged in 50 mL batches and then washed and sonicated with H₂O.The isolated solid was then dried under high vacuum to give Compound 226as a colourless solid (80 mg, 91%).

¹H NMR (500 MHz, DMSO-d₆) δ 12.04 (s, 27H, COOH), 8.00 (s, 3H, ArH),7.93 (d, J = 8.5 Hz, 3H, ArH), 7.78 (s, 3H, NH), 7.73 (s, 3H, NH), 7.54(d, J = 8.5 Hz, 3H, ArH), 7.41 (s, 3H, NH), 7.27 (s, 9H, NH), 6.54 (s,3H, NH), 6.44 (s, 3H, NH), 4.36 (s, 12H, benzylic CH₂), 2.42 (s, 9H,methyl CH₃), 2.35 (s, 9H, methyl CH₃), 2.25 - 1.71 (m, 144H, dendrimerCH₂).

¹³C NMR (126 MHz, DMSO-d₆) δ 174.9, 172.8, 166.0, 156.6, 155.2, 135.9,129.4, 128.8, 124.8, 123.7, 119.7, 67.5, 57.9, 56.8, 31.2, 30.8, 29.5,28.5, 16.3.

Compound 227

A Schlenk flask equipped with a magnetic stirrer was charged withCompound 108 (200 mg, 0.04 mmol, 1.0 eq), DMAP (14.5 mg, 0.12 mmol, 3.0eq) and n-octyl glucoside (23.2 mg, 0.08 mmol, 2.0 eq) dissolved inanhydrous CH₂Cl₂ (40 mL) and then warmed to 34° C. A solution of1,3,5-triisocyanatobenzene (12.5 mg, 0.051 mmol) in toluene (ca. 85%purity) was added to the flask and the reaction was left for 16 h. Thesolvent was removed under vacuum and the crude product was purified byreverse phase MPLC on a C18 SNAP Ultra 60 g cartridge (eluting withAcetone:Water 70:30 to 100:0 over 12CV) giving Compound 227 as a whitesolid (67 mg, 0.012 mmol, 32%).

¹H NMR (500 MHz, CD₃OD) δ 8.11 (d, J = 8.6 Hz, 3H, ArH), 7.76 - 7.69 (m,6H, ArH), 7.14 (s, 3H, ArH), 4.43 (s, 6H, benzylic CH₂), 4.29 (s, 6H,benzylic CH₂), 2.80 - 2.70 (m, 6H, ethyl CH₂), 2.31 - 1.86 (m, 144H,dendrimer CH₂), 1.43 (s, 243H, dendrimer CH₃), 1.21 (t, J = 7.4 Hz, 9H,ethyl CH₃).

HRMS (ESI⁺) calculated for C₂₇₉H₄₅₀N₂₄O₇₂Na₄ ⁴⁺: requires 1345.7981,found [M+4Na]⁴⁺: 1345.7985.

Compound 228

A 50 mL flask, equipped with a magnetic stirrer and a tapped gasadapter, was charged with Compound 13 (89 mg, 0.045 mmol). Compound 13was placed under vacuum for 5 min, then placed under nitrogen. TheCompound 13 was dissolved in dry pyridine (20 mL, 0.002 M), and thesolution heated to 40° C. A solution of Compound 103 (16 mg, 0.050 mmol)in dry dichloromethane (1.0 mL, 0.050 M) was added at a rate of 0.1mL/h. The reaction stirred for a further 12 h at 40° C. before beingconcentrated under vacuum. The residue was purified by reverse phaseflash chromatography on a 60 g SNAP Ultra C18 cartridge elution (1 CV60% acetone/ water, 10 CV 60- 100% acetone/ water, 6 CV 100% acetone).

¹H NMR (500 MHz, CD₃OD) δ 7.99 (d, J = 8.5, 3H, ArH), 7.95 (d, J = 2.1,3H, ArH), 7.62 (s, 3H, RNHCO), 7.53 (dd, J = 8.6, 2.1, 3H, ArH), 4.48(s, 6H, BnH), 4.41 (s, 6H, BnH), 2.85 (q, J = 7.5, 6H, CH₂CH₃), 2.75 (q,J = 7.5, 6H, CH₂CH₃), 2.28 (t, J = 8.0, 18H,CCH₂CH₂), 2.10 (t, J = 8.0,18H, CCH₂CH₂), 1.45 (s, 81H, ^(t)BuH), 1.21 (t, J = 7.4, 18H, CH₂CH₃).

¹³C NMR (126 MHz, CD₃OD) δ 208.96 (C), 173.18 (C), 168.38 (C), 156.81(C), 155.83 (C), 143.16 (C), 143.10 (C), 134.95 (C), 132.69 (C), 132.24(C), 129.85 (C), 127.95 (C), 123.89 (CH), 123.46 (CH), 120.73 (CH),80.34 (C), 58.34(C), 37.39 (CH₂), 29.42 (CH₂), 29.19 (CH₂), 26.98 (CH₃),22.25 (CH₂), 15.25 (CH₃), 15.22 (CH₃).

HRMS (Nanospray) calculated for C₁₂₃H₁₈₅N₁₅O₂₇. requires 1152.6793,found [M+2H]²⁺: 1152.6798.

Compound 229 — Receptor 9

A 10 mL flask, equipped with a magnetic stirrer, was charged withCompound 228 (27 mg, 0.012 mmol). Compound 228 was dissolved indichloromethane (2.9 mL, 0.004 M) before trifluoroacetic acid was added.The reaction stirred for 12 h at rt. The reaction mixture was drippedinto rapidly stirring water (150 mL) which resulted in the formation ofa white precipitate. The precipitate was centrifuged out and dissolvedin acetone. After being transferred to a round bottomed flask, theacetone was removed under vacuum, and the white solid azeotroped withtoluene.

¹H NMR (600 MHz, D₂O) δ 7.86 (s, 3H, ArH), 7.76 (d, J= 8.5, 3H, ArH),7.52 (d, J= 8.5, 3H, ArH), 4.44 (s, 12H, BnH), 2.84- 2.64 (m, 12H,CH₂CH₃), 2.29- 2.16 (m, 18H, CCH₂CH₂), 2.15-2.01 (m, 18H, CCH₂CH₂),1.24- 1.10 (m, 18H, CH₂CH₃).

¹³C NMR (126 MHz, DMSO- d₆) δ 174.50 (C), 166.09 (C), 155.49 (C), 154.56(C), 142.30 (C), 134.21 (C), 132.81 (C), 128.34 (C), 128.20 (C), 123.05(CH), 122.59 (CH), 119.33 (CH), 57.10 (C), 37.04 (CH₂), 29.06 (CH₂),28.22 (CH₂), 21.04 (CH₂), 16.34 (CH₃).

Compound 230

Compound 11 (1.384 g, 2.409 mmol) and Compound 5a (0.200 g, 0.611 mmol)were placed in a round bottomed flask under nitrogen and dissolved inanhydrous DMF (21 mL). To this solution was added dry pyridine (0.147mL, 1.833 mmol) then heated to 30° C. for 100 hours. The solvent wasevaporated under vacuum to give a gum which was purified by columnchromatography by pre-adsorbing onto silica gel (20 g) by dissolving thecrude in a mixture of dichloromethane and methanol and removing thesolvent under vacuum to give a free-flowing powder. This pre-adsorbedmaterial was loaded into an empty cartridge and put in line with a 100 gSNAP HP Sil cartridge and eluting with a gradient of dichloromethanewith increasing concentration of methanol gave recovered Compound 11(367 mg), and Compound 230 (1.171 g, 93%).

¹H NMR: (400 MHz, (CD₃OD): δ m.br. 7.95-7.20 (33H, ArH), s. 4.40 (6H,ArCH₂NH), m. 4.30-4.10 (9H, FmocH), m. 3.71-3.60 (42H, CH₂CH₂O, andOCH₂CH₂NH₂), m. 3.33 (6H, OCH₂CH₂N₃), m.br. 2.79 (6H, ArCH₂CH₃), s.br.1.19 (9H, ArCH₂CH₃).

Compound 231

To a stirred suspension of Compound 230 (1.171 g, 0.571 mmol) in DCM (5mL) at room temperature was added distilled DBU (0.426 mL, 2.854 mmol).After 10 minutes the reaction mixture became a clear solution and wasstirred for a total of 2 hours before loading directly onto a flashchromatography column 50 g SNAP KP Sil cartridge that was equilibriatedwith dichloromethane and then eluting with a gradient of dichloromethanewith increasing concentration of methanol to give the desired productcontaminated with the fluorenyl by-product. Partial evaporation of theproduct-containing fractions gave a thick white slurry where the solidwas separated by centrifuge. The wet solid was resuspended in methanoland centrifuged again and the solid dried to give Compound 231 as anoff-white solid (0.344 g, 43%).

¹H NMR (400 MHz, CDCl3/methanol-d₄) δ m. 7.12-6.85 (9H, ArH), s. 4.32(6H, ArCH₂NH), m. 3.50-3.40 (42H, CH₂CH₂O, and OCH₂CH₂N₃), t. 3.20 (6H,OCH₂CH₂N₃), m.br. 2.66 (6H, CH₂), s.br. 1.09 (9H, CH₃).

Compound 232

Dissolved Compound 231 (226 mg, 163 mmol) in dry DMSO (1 mL) and dilutedwith dry pyridine (70 mL). Stirred under nitrogen at 40° C. and added asolution of Compound 5a (60 mg, 183 mmol) in dry DCM (2 mL) by syringepump over 6 hours, then left stirring at 40° C. for a further 60 hours.Evaporation of the solvent on a rotary evaporator gave an orangecoloured gum which was dissolved in methanol containing a little water.This was loaded onto a Biotage 120 g reverse phase column and elutedwith a water/methanol gradient. The compound which eluted at about 80%methanol was collected, evaporated, redissolved in methanol,concentrated by boiling off the solvent and the hot solution (~3 mL) wasleft to cool overnight to give the product (Compound 232) as clumps ofwhite crystals (83 mg, 30%).

¹H NMR (400 MHz, D₂O) δ s. 8.09 (3H, ArH), d. 7.98 (3H, ArH), d. 7.58(3H, ArH), m. 4.41-4.45 (12H, ArCH₂NH), m. 3.5-3.7 (42H, OCH₂CH₂), m.2.7-2.9 (12H, ArCH₂CH₃), m. 1.20 (18H, ArCH₂CH₃).

HRMS: (ESI⁺) calculated for C₃₁H₁₁₃N₂₄O₁₃ ²⁺ [M+2H]²⁺ 856.4449, found:856.4470.

Compound 233 — Receptor 10

Compound 232 (22 mg, 0.013 mmol) was dissolved in a mixture of warmmethanol (2 mL) and water (0.1 mL) and to this was addedtriphenylphosphine (26 mg, 0.099 mmol) under an atmosphere of nitrogen.The reaction mixture was then heated for 16 hours at 60° C. beforeallowing to cool to room temperature. The cloudy mixture was dilutedwith more methanol and water, followed by 40 µL of 1 M aqueoushydrochloric (until it was acidic). Added more water then extracted it acouple of times with DCM to remove the triphenylphosphine basedcompounds. The slightly cloudy aqueous layer was passed through abond-elut (500 mg) several times until the eluant emerged clear and tlcshowed that no product was passing through. Then eluted with water 6 mL,then 2 x 4 mL of 25% MeOH in water, then 4 x 4 mL of 50%, 2 x 4 mL of75% MeOH in water. Tlc showed that the product had eluted in the 25% to75% methanol fractions which were combined and evaporated before beingredissolved in water and freeze-dried to give Compound 233 (19 mg, 89%)as an off-white solid.

¹H NMR (400 MHz, D20) δ s. 7.88 (3H, ArH), d. 7.71 (3H, ArH), d. 7.43(3H, ArH), s.br. 4.20 (12H, ArCH₂NH), m. 3.4-3.6 (42H, OCH₂CH₂, andOCH₂CH₂NH₂), t. 3.00 (6H, OCH₂CH₂NH₂), m. 2.48 (12H, ArCH₂CH₃), m. 0.96(18H, ArCH₂CH₃).

HRMS: (MALDI⁺) calculated for C₈₁H₁₂₀N₁₈O₁₈Na⁺ [M+Na]⁺ 1655.8920, found:1655.8932.

Hexa-carboxylate Macrocycle (Compound H8 - Receptor 11 )

N-(4,5-dimethyl-2-nitrophenyl)acetamide (Compound H2)

4,5-dimethyl-2-nitroaniline (Compound H1, 2 g, 12 mmol) was suspended inglacial acetic acid (24 mL) and heated to 90° C. Acetic anhydride (1.2mL, 13 mmol) was added and the mixture stirred at reflux for 2 hours.The reaction mixture was cooled to room temperature and poured intowater (300 mL). The yellow precipitate was filtered, washed with waterand recrystallized from ethanol to yield Compound H2 (2.4 g, 11.6 mmol,97%) as a yellow crystalline solid.

¹H NMR: (400 MHz, (CDCl₃): δ 2.25 (s, 6H, C(9, 10)H₃), 2.32 (s, 3H,C(1)H₃), 7.93(s, 1H, C(8)H), 8.51 (s, 1H, C(5)H), 10.26 (br s, 1H, NH).

¹³C NMR: (100 MHz, (CDCl₃): δ 19.1 (C10), 20.5 (C9), 25.6 (C1), 122.6(C8), 125.9 (C5), 132.3 (C7), 132.7 (C6), 134.1 (C3), 146.8 (C4), 168.9(C(2)O); v _(max) 3341, 2987, 2901, 1708, 1695, 1576, 1323, 1151, 759cm⁻¹.

HRMS: (ESI⁺) Found [M+Na]⁺: 231.0745.

4-Aminonitrophthalic Acid (Compound H3)

Under an inert N₂ atmosphere, Compound H2 (2 g, 9.6 mmol) and KMnO₄ (6g, 37.9 mmol)) was suspended in water (50 mL) and stirred at reflux for3 days. Additional KMnO₄ (3 g, 19 mmol) was added halfway through thereaction time. The resultant brown precipitate was filtered hot andwashed with water. The yellow filtrate was acidified to pH 3 with 1 MHCI, extracted with EtOAc (3 × 100 mL), washed with brine (100 mL) anddried (MgSO₄). The solvent was removed under vacuum to yield Compound H3(0.77 g, 2.88 mmol, 60%) as a yellow orange solid.

¹H NMR: (400 MHz, (CDCl₃): δ 7.25 (s, 1H, C(6)H), 7.63 (br s, 2H, NH₂),8.70 (s, 1H, C(3)H), 11.50 (br s, 2H, C(7, 8)O₂H).

¹³C NMR: (100 MHz, (CDCl₃): δ 117.7 (C4), 118.7 (C6), 129.3 (C3), 132.6(C5), 141.5 (C2), 146.9 (C1), 167.8 (C7), 169.0 (C8); v _(max) 3486,3364, 2972, 2901, 1712, 1681, 1626, 1502, 1252, 1057, 882 cm⁻¹.

LRMS: (El) Found [M]⁺: 226.1.

Dimethyl 4-amino-5-nitrophthalate (Compound H4)

Compound H3 (0.8 g, 3.5 mmol) was dissolved in MeOH (30 mL) andconcentrated H₂SO₄ (0.5 mL) added. The reaction mixture was stirred atreflux for 3 hours and then the solvent was removed under vacuum. Theresidue was dissolved in EtOAc (60 mL), washed with 5% NaHCO₃ (60 mL),brine (60 mL) and dried (MgSO₄). The solvent was removed under vacuumand the crude solid purified by flash column chromatography (100%CH₂Cl₂) to yield Compound H4 (0.72 g, 2.8 mmol, 80%) as an orange solid.

¹H NMR: (400 MHz, (CDCl₃): δ 3.88 (s, 3H, C(10)H₃), 3.92 (s, 3H,C(9)H₃), 6.91 (s, 1H, C(6)H), 7.26 (br s, 2H, NH₂), 8.74 (s, 1H, C(3)H).

¹³C NMR: (100 MHz, (CDCl₃): δ 52.5 (C9), 53.1 (C10), 116.9 (C4), 118.3(C6), 129.7 (C3), 131.2 (C5), 140.7 (C2), 146.6 (C1), 164.7 (C7), 168.0(C8); v _(max) 3486, 3342, 2987, 2901, 1736, 1697, 1621, 1502, 1435,1339, 1250, 1027, 762 cm⁻¹.

LRMS: (ESI⁺) Found [M+Na]⁺: 277.1.

Dimethyl 4-amino-5-nitrophthalate (Compound H5)

Under an inert N₂ atmosphere, a solution of Compound H4 (0.1 g, 0.39mmol) in MeOH (10 mL) was added to Pd/C (10 mg). The reaction vessel wasthen purged with hydrogen (1 atm) and the reaction mixture stirred atroom temperature for 1 hour. After which, the reaction mixture wasfiltered through celite and washed with CH₂Cl₂, and the filtrateconcentrated under vacuum. The crude product was then purified by flashcolumn chromatography (5% MeOH:CH₂Cl₂) to afford Compound H5 (81 mg,0.36 mmol, 93%) as light brown solid.

¹H NMR: (400 MHz, (CDCl₃): δ 3.84 (s, 6H, C(5)H₃), 7.02 (s, 2H, C(2)H).

¹³C NMR: (100 MHz, (CDCl₃): δ 52.4 (C5), 116.6 (C2), 124.1 (C3), 1136.6(C1), 168.4 (C4).

HRMS: (ESI⁺) Found [M+Na]⁺: 247.0685.

Hexa-ester Amino Half Receptor (Compound H6)

Under an inert N₂ atmosphere, Compound H5 (80 mg, 0.36 mmol) wasdissolved in dry pyridine (10 mL) and heated to 40° C. A solution of TEBisocyanate (Compound 103, 20 mg, 0.06 mmol) in dry CH₂Cl₂ (2 mL) wasadded over 1 hour and the reaction stirred at 40° C. for 16 hours. Thereaction mixture was concentrated under vacuum and residual pyridineco-evaporated with toluene (3 × 30 mL). The crude product was thensuspended in CH₂Cl₂, filtered and air dried to afford Compound H6 (49mg, 0.049 mmol, 82%) as a light brown solid.

¹H NMR: (400 MHz, ((CD₃)₂SO): δ 1.19 (t, J = 7.2 Hz, 9H, C(1)E₃), 2.79(br q, 6H, C(2)H₂), 3.72, 3.73 (s, 2 × 9H, C(14, 16)H₃), 4.36 (s, 6H,C(5)H₂), 5.87 (br s, 6H, NH₂), 6.51 (br t, 3H, NHC(5)), 6.84 (s, 3H,C(11)H), 8.13 (s, 3H, C(8)H), 8.60 (s, 3H, NH).

¹³C NMR: (100 MHz, ((CD₃)₂SO): δ 16.9 (C1), 22.8 (C2), 37.7 (C5), 52.2,52.5 (C14 and C16), 114.0 (C11), 117.3 (C9), 121.7 (C8), 126.9 (C7),128.6 (C10), 133.2 (C4), 142.2 (C12), 143.4 (C4), 155.5 (C6), 167.3,169.2 (C9).

HRMS: (ESI⁺) Found [M+H]⁺: 1000.4041.

Hexa-ester Hexa-urea Macrocycle (Compound H7)

Method A: Under an inert N₂ atmosphere, Compound H5 (40 mg, 0.18 mmol)was dissolved in dry pyridine (100 mL) and heated to 40° C. A solutionof TEB isocyanate (Compound 103, 38 mg, 0.12 mmol) in dry CH₂Cl₂ (2 mL)was added over 1 hour and the reaction stirred at 40° C. for 16 hours.The reaction mixture was concentrated under vacuum and residual pyridineco-evaporated with toluene (3 × 30 mL). The crude product was thenpurified by reverse phase HPLC (100% water → 100% acetonitrile) toafford Compound H7 (9.5 mg, 0.007 mmol, 12%) as a white solid.

Method B: Under an inert N₂ atmosphere, Compound H6 (20 mg, 0.02 mmol)was dissolved in dry pyridine (20 mL) and heated to 40° C. A solution ofTEB isocyanate (Compound 103, 7.8 mg, 0.024 mmol) in dry CH₂Cl₂ (2 mL)was added and the reaction stirred at 40° C. for 16 hours. The reactionmixture was concentrated under vacuum and residual pyridineco-evaporated with toluene (3 × 30 mL). The crude product was thenpurified by reverse phase HPLC (100% water → 100% acetonitrile) toafford Compound H7 (8 mg, 0.006 mmol, 31%) as a white solid.

¹H NMR: (400 MHz, (CD₃OD): δ 1.22 (t, J = 7.4 Hz, 18H, C(1)H₃), 2.79 (q,J = 7.4 Hz, 12H, C(2)H₂), 3.86 (s, 18H, C(11)H₃), 4.42 (s, 12H, C(5)H₂),8.34 (s, 6H, C(8)H).

¹³C NMR: (100 MHz, (CD₃OD): δ 15.3 (C1), 22.4 (C2), 37.5 (C5), 51.5(C11), 121.6 (C8), 126.3 (C7), 131.6 (C3), 131.9 (C9), 143.2 (C4), 155.6(C6), 168.1 (C10).

HRMS: (ESI⁺) Found [M+H]⁺: 1327.5635.

Hexa-carboxylate Hexa-urea Macrocycle (Compound H8 - Receptor 11)

Compound H7 (8 mg, 0.006 mmol) was dissolved in MeOH (4 mL) and thenNaOH (5 M, 1 mL) added dropwise. The solution was stirred at 40° C. for1 hour and the reaction diluted with water (5 mL). The MeOH was removedunder vacuum and the aqueous solution neutralised to pH 7.4 with acidicion exchange resin, filtered and freeze dried to afford Compound H8 (7.8mg, 0.0056 mmol, 93%) as a white solid.

¹H NMR: (600 MHz, D₂O): δ 1.19 (t, J = 7.4 Hz, 18H, C(1)E₃), 2.75 (br q,12H, C(2)H₂), 4.47 (s, 12H, C(5)H₂), 7.73 (s, 6H, C(8)H).

¹³C NMR: (100 MHz, D₂O): δ 15.4 (C1), 22.5 (C2), 37.6 (C5), 124.6 (C8),128.3 (C7), 131.8 (C3), 134.9 (C9), 143.3 (C4), 157.4 (C6), 176.4 (C10).

Compound 234

Compound 15 (2.000 g, 3.363 mmol) was dissolved in THF (145.0 mL) andmixed with triphenylphosphine (0.838 g, 3.195 mmol). The reaction wasstirred at room temperature for 24 h. Water (75 mL) was added and thereaction heated at 50° C. for 3 h. After cooling the reaction mixturewas diluted with water and extracted with EtOAc (3 × 100 mL). Combinedorganic layers were concentrated in vacuo and the crude residue purifiedby reverse phase MPLC on a C18 SNAP Ultra 120 g cartridge eluting (40%acetone:water to 60% acetone:water) to give Compound 234 as a whitesolid (1.510 g, 2.655 mmol, 79%).

¹H NMR (400 MHz, CD₃OD) δ 4.57 (s, 1H, NH), 3.86 (dt, J = 5.8, 4.0 Hz,1H, NCH₂), 3.73 (dd, J = 11.2, 5.8 Hz, 1H, NCH₂), 3.59 (dd, J = 10.6,6.1 Hz, 1H, NCH₂), 3.41-3.32 (m, 2H, CHN₃, CHNH₂), 3.13 (dd, J = 10.6,4.1 Hz, 1H NCH₂), 2.22 (m, 6H, CH₂C(O)), 1.95 (m, 6H, CCH₂), 1.45 (s,27H, C(CH₃)₃).

¹³C NMR (100 MHz, CD₃OD) δ 174.9 (s, CCO₂C), 157.8 (s, NC(O)N), 81.7 (s,CO₂C(CH₃)₃), 67.2 (s, CNH₂), 58.3 (CN₃), 56.8 (s, CH₂N), 52.5 (CH₂N),31.2 (s, CH₂C(O)), 30.8 (s, CCH₂), 28.4 (s, CO₂C(CH₃)₃).

Compound 235

Compound 234 (67 mg, 0.118 mmol) and Compound 103 (11 mg, 0.034 mmol)were dissolved in DCM (1 mL) and stirred together for 18 h. The solventwas removed in vacuo and the residue purified by reverse phase MPLC on aC18 SNAP Ultra 120 g cartridge eluting 70% acetone:water to 100%acetone:water to give Compound 235 as a white solid (60 mg, 0.030 mmol,89%).

¹H NMR (400 MHz, CD₃OD) δ 4.39 (s, 6H, ArCH₂NH), 4.24 (dt, J = 6.7, 4.7Hz, 3H, CHNH), 4.05 (dt, J = 6.0, 4.5 Hz, 3H, NCH₂), 3.68 (dd, J = 10.8,6.7 Hz, 3H, NCH₂), 3.61 (dd, J = 11.3, 6.0 Hz, 3H, NCH₂), 3.32 (m, 3H,CHN₃), 3.17 (dd, J = 10.8, 4.6 Hz, 3H NCH₂), 2.78 (q, J = 7.4 Hz, 6H,ArCH₂CH₃), 2.21 (dd, J = 9.3, 6.6 Hz, 18H, CH₂C(O)), 1.94 (dd, J = 9.3,6.6 Hz, 18H, CCH₂), 1.44 (s, 81H, C(CH₃)₃), 1.18 (t, J = 7.4 Hz, 9H,ArCH₂CH₃).

¹³C NMR (100 MHz, CD₃OD) δ 174.9 (s, CCO₂C), 157.8, 152.4 (s, NC(O)N),137.5, 133.7 (Ar), 81.8 (s, CO₂C(CH₃)₃), 69.1 (s, CHNH), 61.5 (CN₃),58.5 (s, CH₂N), 51.0 (CH₂N), 39.3 (ArCH₂NH), 31.2 (s, CH₂C(O)), 30.9 (s,CCH₂), 28.4 (s, CO₂C(CH₃)₃), 22.2 (s, ArCH₂CH₃), 15.6 (s, ArCH₂CH₃).

Compound 236

To a solution of Compound 235 (100 mg, 0.049 mmol) in MeOH (15 mL) wasadded a slurry of Pd/C (30 mg) in DCM. The reaction was placed under ahydrogen atmosphere and left to stir overnight. Filtration throughCelite™ and concentration of the filtrate gave a white solid (91 mg,0.047 mmol, 96%).

¹H NMR (400 MHz, CD₃OD) δ 4.46-4.33 (m, 6H, ArCH₂NH), 4.25 (d, J = 6.9Hz, 3H, CHNH), 3.88-3.74 (m, 6H, NCH₂), 3.60 (d, J = 6.4 Hz, 3H, NCH₂),3.33-3.27 (m, 3H, CHNH₂), 3.20 (dd, J = 10.4, 7.0z Hz, 3H, NCH₂), 2.78(q, J = 8.2 Hz, 6H, ArCH₂CH₃), 2.30-2.14 (m, 18H, CH₂C(O)), 2.04-1.85(m, 18H, CCH₂), 1.44 (s, 81H, C(CH₃)₃), 1.19 (t, J = 7.4 Hz, 9H,ArCH₂CH₃).

¹³C NMR (100 MHz, CD₃OD) δ 174.8 (s, CCO₂C), 157.9, 151.5 (s, NC(O)N),137.2, 132.0 (Ar), 81.7 (s, CO₂C(CH₃)₃), 68.1 (s, CHNH), 67.5 (CNH₂),58.2 (s, CH₂N), 54.8 (CH₂N), 43.6 (ArCH₂NH), 31.2 (s, CH₂C(O)), 30.8 (s,CCH₂), 28.4 (s, CO₂C(CH₃)₃), 22.3 (s, ArCH₂CH₃), 16.9 (s, ArCH₂CH₃).

Compound 237

A mixture of Compound 236 (110 mg, 0.056 mmol), n-octyl glucoside (33mg, 0.112 mmol) and DMAP (21 mg, 0.168 mmol) were azeotroped to drynesswith toluene in a two-necked flask and then placed under N₂. The residuewas then dissolved in DCM (110 mL) and cooled to 0° C. A solution of TEBNCO (Compound 103, 18 mg, 0.056 mmol) in DCM (20 mL) was added. Thereaction mixture was heated to 35° C. for 16 h. Solvent was removedunder reduced pressure, and the crude product purified by reverse phaseMPLC on a C18 SNAP Ultra 60 g cartridge eluting 70% acetone:water to100% acetone:water (46 mg, 0.020 mmol, 36%).

¹H NMR (500 MHz, CD₃OD) δ 4.79-4.33 (m, 6H, ArCH₂NH), 4.31-4.10 (m, 6H,ArCH₂NH), 3.97-3.80 (m, 6H, CHNH), 3.73-3.37 (m, 9H, NCH₂), 2.96-2.56(m, 9H, NCH₂, ArCH₂CH₃), 2.35-2.20 (m, 24H, ArCH₂CH₃, CH₂C(O)),2.10-1.87 (m, 18H, CCH₂), 1.48-1.40 (m, 81H, C(CH₃)₃), 1.24-1.10 (m,18H, ArCH₂CH₃).

¹³C NMR (125 MHz, (CD₃)₂SO) δ 172.5 (s, CCO₂C), 157.6, 155.4 (s,NC(O)N), 141.6, 133.9 (Ar), 79.7 (s, CO₂C(CH₃)₃), 56.2 (NHC(CH₂)₃), 52.7(s, CHNH), 50.2 (s, CH₂N), 36.6 (ArCH₂NH), 29.4 (s, CH₂C(O)), 29.3 (s,CCH₂), 27.8 (s, CO₂C(CH₃)₃), 22.0 (s, ArCH₂CH₃), 16.3 (s, ArCH₂CH₃).

MS: (ESI⁺) calculated for C₁₁₇H₁₉₅N₁₈O₂₇ ²⁺. 1141.7180, found [M+2H]²⁺:1141.7196

Compound 238 - Receptor 12

Compound 237 (8 mg, 3.504 µmol) dissolved in TFA (1.3 mL), and heated to30° C. with stirring for 20 hours. The reaction mixture was allowed tocool, whereupon pentane (25 mL) was added. The resultant suspension wascentrifuged, and the supernatant removed. The residual oil was dissolvedin 0.2 _(M) aq. NaHCO₃ solution (3.5 mL) and this solution was desaltedby 20 mL column of G-25 sephadex. The resultant solution was freezedried to give Compound 238 as a white solid (6.5 mg, 3.290 mmol, 94%).

¹H NMR (500 MHz, D₂O) δ 4.66-4.50 (m, 3H, ArCH₂NH), 4.27-4.03 (m, 9H,ArCH₂NH), 4.02-3.82 (m, 3H, CHNH), 3.79-3.34 (m, 12H, CHNH, NCH₂),3.09-2.75 (m, 6H, ArCH₂CH₃), 2.75-2.27 (m, 9H, NCH₂, ArCH₂CH₃),2.26-2.13 (m, 18H, CH₂C(O)), 2.03-1.89 (m, 18H, CCH₂), 1.23-1.09 (m,18H, ArCH₂CH₃).

¹³C NMR (125 MHz, D₂O) δ 183.4 (s, CCO₂), 160.2, 159.7, 159.3(NHC(O)NH), 157.6 (s, NHC(O)N), 144.9, 144.2, 144.1 (Ar), 58.1(NHC(CH₂)₃), 58.0, 57.9 (s, CHNH), 49.1, 48.4, 47.7 (s, CH₂N), 39.8,38.8, 37.6 (ArCH₂NH), 32.2, 32.1 (s, CH₂C(O)), 31.7 (s, CCH₂), 23.5,22.7, 22.1 (s, ArCH₂CH₃), 16.1, 15.9, 15.5 (s, ArCH₂CH₃).

MS: (ESI⁺) calculated for C₁₁₇H₁₉₅N₁₈O₂₇ ²⁺. 1141.7180, found [M+2H]²⁺:1141.7196

Compound 239

A 1 L flask, with a side-arm gas adaptor, equipped with a magneticstirrer was dried under vacuum using a heat-gun. The flask was cooled toRT and charged with G2 amine (Compound 82, 2.50 g, 1.74 mmol) underflowing nitrogen. THF (100 mL) and Et₃N (0.25 mL, 1.80 mmol) were added,and the flask cooled to 0° C. with an ice bath. A solution oftriphosgene (0.26 g, 1.00 mmol) in THF (25 mL) was added dropwise overthe course of 20 min. After 3 h, the solvent was removed under vacuumand the resultant residue taken up in chloroform (50 mL) and washed withwater (50 mL). The organic layer was dried (MgSO₄) and concentratedunder vacuum to give Compound 239 as a white foam (2.50 g, 1.71 mmol,98%).

¹H NMR (400 MHz, toluene-d₈) δ 2.26 (m, 18H, CH₂), 2.07 (m, 30H, CH₂),1.40 (s, 81H, CH₃).

¹³C NMR (100 MHz, toluene-d₈) δ 173.3 (CCO₂C), 172.4 (CONH), 123.7(N=C=O), 80.4 (C(CH₃)₃), 58.2 (NHC(CH₂)₃), 53.7 (C(NCO)), 30.7 (CH₂),30.5 (CH₂), 28.7 (CH₂), 28.5 (CH₃).

Compound 240

A Schlenk flash was charged with (3S,4S)-pyrrolidine-3,4-diol (185.0 mg,1.790 mmol), G2 NCO (2.500 g, 2.000 mmol) and anhydrous DMF (100 mL)under N₂. The solution was left to stir for 16 hours, then poured into5% aq. LiCl (700 mL) and extracted with EtOAc (300 mL). The organiclayer was separated, dried and concentrated to a gummy solid (2.760 g,1.759 mmol, 88%).

¹H NMR (400 MHz, CDCl₃) δ 6.24 (s, 3H, C(O)NH), 6.13 (2H, s, OH), 4.19(s, 1H, NC(O)NH), 3.63 (dd, J = 11.5, 3.7 Hz, 2H, CHOH), 3.54-3.37 (m,4H, NCH₂), 2.17 (dd, J = 10.3, 6.4 Hz, 24H, CH₂C(O)), 1.92 (dd, J = 9.9,6.4 Hz, 24H, CCH₂), 1.41 (s, 81H, C(CH₃)₃).

¹³C NMR (100 MHz, CDCl₃) δ 173.4 (C(O)NH), 172.9 (CCO₂C), 162.6 (s,NC(O)N), 80.8 (s, CO₂C(CH₃)₃), 80.7 (s, COH), 57.4 (s, CH₂N), 30.1 (s,CH₂C(O)), 30.0 (s, CH₂C(O)NH), 29.9 (s, CCH₂), 29.9 (CCH₂), 28.2 (s,CO₂C(CH₃)₃).

Compound 241

Compound 240 (1.851 g 1.180 mmol) was dissolved in DCM (13.9 mL), andtriethylamine (0.66 mL, 4.719 mmol) was added. The reaction was cooledto 0° C. and mesyl chloride (0.20 mL, 2.595 mmol) was added dropwise.The reaction was stirred at room temperature for 16 h, before beingwashed with 5% aq. KHSO₄, sat. aq. NaHCO₃ and brine. The organic layerwas concentrated in vacuo to give Compound 241 as a white foam (1.350 g,0.783 mmol, 66%).

¹H NMR (400 MHz, CDCl₃) δ 6.95 (s, 1H, NC(O)NH), 6.17 (s, 3H, C(O)NH),5.32-5.13 (m, 2H, CHOSO₂), 3.94-3.56 (m, 4H, NCH₂), 3.13 (s, J = 1.6 Hz,2H, SO₂CH₃), 2.31-2.11 (m, 24H, CH₂C(O)), 2.04-1.86 (m, 24H, CCH₂), 1.41(s, 81H, C(CH₃)₃).

¹³C NMR (100 MHz, CDCl₃) δ 173.3 (C(O)NH), 172.7 (CCO₂C), 156.3 (s,NC(O)N), 80.6 (s, CO₂C(CH₃)₃), 80.1 (s, COSO₂), 57.4 (s, CH₂N), 38.6(SO₂CH₃), 29.9 (s, CH₂C(O)), 29.9 (s, CH₂C(O)NH), 29.8 (s, CCH₂), 29.8(CCH₂), 28.1 (s, CO₂C(CH₃)₃).

Compound 242

NaN₃ (0.131 g, 2.017 mmol) was added to a solution of Compound 241(1.160 g, 0.672 mmol) in DMF (3.4 mL) at 0° C. The reaction was thenheated at 100° C. for 16 h. The reaction was cooled and diluted withEtOAc (30 mL) and washed with water (30 mL), 5% aq. LiCl (2 × 30 mL) andbrine (30 mL). The organic layer was concentrated in vacuo and theresidue purified by reverse phase MPLC on a C18 SNAP Ultra 60 gcartridge eluting (70% acetone:water to 100% acetone:water) to giveCompound 242 as a white solid (788 mg, 0.487 mmol, 72%).

¹H NMR (400 MHz, CD₃OD) δ 4.29-4.18 (m, 2H, CHN₃), 3.84-3.70 (m, 2H,NCH₂), 3.55-3.44 (m, 2H, NCH₂), 2.32-2.19 (m, 24H, CH₂C(O)), 2.06-1.98(m, 24H, CCH₂), 1.52 (s, 81H, C(CH₃)₃).

¹³C NMR (100 MHz, CD₃OD) δ 175.8 (C(O)NH), 174.4 (CCO₂C), 158.1 (s,NC(O)N), 81.7 (s, CO₂C(CH₃)₃), 65.1 (s, CH₂N₃), 62.2 (NC(O)NHC(CH₂)₃),61.5 (NHC(CH₂)₃), 58z.8 (CH₂N), 30.7 (s, CH₂C(O)), 30.7 (s, CH₂C(O)NH),30.5 (s, CCH₂), 30.5 (CCH₂), 28.4 (s, CO₂C(CH₃)₃).

Compound 243

Compund 241 (300 mg, 0.185 mmol) was dissolved in THF (8.0 mL).Triphenylphosphine (46 mg, 0.176 mmol) was added and the reactionstirred for 18 h. Water (4.0 mL) was added and the reaction heated at50° C. for 6 h. The reaction mixture was concentrated in vacuo and theresidue purified by reverse phase MPLC on a C18 SNAP Ultra 60 gcartridge eluting (75% acetone:water to 100% acetone:water) to give aCompound 243 as a white solid (187 mg, 0.117 mmol, 66%).

¹H NMR (400 MHz, CD₃OD) δ 3.92-3.72 (m, J = 5.8, 4.0 Hz, 3H, NCH₂), 3.61(dd, J = 10.7, 6.0 Hz, 1H, CHN₃), 3.41-3.32 (m, 1H, CHNH₂), 3.24-3.12(m, 1H NCH₂), 2.23-2.16 (m, 24H, CH₂C(O)), 1.99-1.91 (m, 24H, CCH₂),1.45 (s, 81H, C(CH₃)₃).

¹³C NMR (100 MHz, CDCl₃) δ 173.3 (C(O)NH), 172.7 (CCO₂C), 158.2 (s,NC(O)N), 81.6 (s, CO₂C(CH₃)₃), 58.7 (CNH₂), 58.6 (s, CH₂N), 58.4 (CN₃),33.1 (s, CH₂C(O)), 32.3 (s, CH₂C(O)NH), 30.7 (s, CCH₂), 30.5 (CCH₂),28.4 (s, CO₂C(CH₃)₃).

Compound 244

Compound 7b (80 mg, 0.016 mmol) was azeotroped with toluene to drynessin reaction flask and then redissolved in pyridine (8.0 mL). This washeated to 40° C. and a solution of Compound 5a (6.2 mg, 0.019 mmol) inDCM (0.7 mL) was added by syringe pump over 3 h. The reaction was thencooled to room temperature and stirred for a further 16 h. The reactionwas concentrated under vacuum and the crude residue obtained was thenpurified by reverse phase flash chromatography on a 30 g SNAP Ultra C18cartridge elution (1CV 80% acetone/H₂O, 10 CV 80-95% acetone/H₂O, 2CV100% acetone) to give Comppound 244 as a white solid (55 mg, 0.010 mmol,64%).

¹H NMR (500 MHz, methanol-d₄) δ ^(z)d. 8.12 (3H, J = 2.1 Hz, Ar), dd.7.56 (3H, J = 8.4, 2.1 Hz, Ar), d. 7.41 (3H, J= 8.4 Hz), s. 4.58 (6H,ArCH₂NH), s. 4.45 (6H, ArCH₂NH), s. 3.85 (9H, ArOCH₃), m. 2.82-2.72 (6H,ArCH₂CH₃), m. 2.35-1.89 (144H, NHCH₂CH₂C(O)), s. 1.43 (243H,CO₂C(CH₃)₃), t. 1.20 (9H, J = 7.3 Hz, ArCH₂CH₃).

¹³C NMR (125 MHz, methanol-d₄) δ 175.5 (CONH), 174.4 (CO₂C(CH₃)₃), 171.1(ArCONHR), 81.6 (CO₂C(CH₃)₃), 59.3 (ArOCH₃), 58.7 (C(CH₂CH₂CO₂)₃), 54.6(C(CH₂CH₂CONH)₃), 44.2, 43.7 (ArCH₂NHC(O)NH), 32.2 (CH₂CH₂CONH,CH₂CH₂CONH), 30.7 (CH₂CH₂CO₂C(CH₃)₃), 30.5, (CH₂CH₂CO₂C(CH₃)₃), 28.5(CO₂C(CH₃)₃), 16.9 (ArCH₂CH₃).

MS: (ESI⁺) calculated for C₂₈₂H₄₅₆N₂₄O₇₅ ³⁺: 1795.0979, found [M+3H]³⁺:1795.0938.

Compound 245 - Receptor 13

Compound 244 (8.5 mg, 0.002 mmol) was dissolved in DCM (0.5 mL) andformic acid (0.5 mL, 13.3 mmol) was added. After 24 h the reactionmixture was added dropwise into stirring water (20 mL) and the whiteprecipitate collected by centrifuging. The supernatant was decanted off,and the white solid was neutralised using 10 mM NaOH solution to pH 7.The resulting solution was desalted by 10 mL column of G-25 sephadex.The resultant solution was freeze dried to give a white solid (1 mg,0.32 µmol, 16%).

¹H NMR (500 MHz, CD₂Cl₂/formic acid-d₂, 1:1) δ ¹H NMR (500 MHz,methanol-d₄) δ s. 8.18 (3H, Ar), s. 7.50 (6H, Ar), s. 4.48 (6H,ArCH₂NH), s. 4.41 (6H, ArCH₂NH), s. 3.83 (9H, ArOCH₃), m. 2.69-2.60 (6H,ArCH₂CH₃), m. 2.45-1.81 (144H, NHCH₂CH₂C(O)), m. 1.16-1.08 (9H,ArCH₂CH₃).

Compound 246

Prepared in a manner analogous to Compound 244 from Compounds 7a (202.0mg, 0.040 mmol) and 5c (11.4 mg, 0.040 mmol). Purified by reverse phaseflash chromatography on a 120 g SNAP Ultra C18 cartridge elution (1CV80% acetone/H₂O, 10 CV 80-97% acetone/H₂O, 4CV 97% acetone) to giveCompound 246 as a white solid (108.0 mg, 50.6%).

¹H NMR (500 MHz, methanol-d₄) δ m. 8.08-7.58 (9H, Ar), m. 4.63-4.30(12H, ArCH₂NH), m. 2.95-2.60 (9H ArCH₃), m. 2.32-1.84 (150H,NHCH₂CH₂C(O), ArCH₂CH₃), s. 1.43 (243H, CO₂C(CH₃)₃), s. 1.29 (9H,ArCH₂CH₃).

Compound 247 - Receptor 14

Compound 246 (108 mg, 0.02 mmol) was dissolved in anhydrous DCM (Vol: 20mL) and TFA (4.8 mL, 60.0 mmol) at RT. The resulting off yellow solutionwas stirred for 12 h at room temperature. Volatiles were removed undervacuum to give a yellow solid. The solid which was purified by reversephase MPLC on a C18 SNAP Ultra 60 g cartridge by loading the sample in1:1 MeOH/H₂O+0.1 % formic acid). The resulting white solid wasneutralised using 100 mM NaOH solution to pH 7 and the resultingsolution concentrated to dryness under vacuum. White crystalline solid(30 mg, 0.008 mmol, 40 %).

¹H NMR (500 MHz, methanol-d₄) δ m. 8.37-7.81 (9H, Ar), m. 4.57-4.22(12H, ArCH₂NH), m. 2.85-1.66 (159H, ArCH₃, NHCH₂CH₂C(O), ArCH₂CH₃), t.1.25 (9H, J = 9.2 Hz, ArCH₂CH₃).

2,3:4,5-bis-O-(1-methylethylidene)-1-O-2-propynyl-L-arabinitol (Compound248)

To a suspension of the sodium hydride (500 mg of 60% by weight in oil,12.9 mmol) in THF (20 mL) was added2,3:4,5-bis-O-(1-methylethylidene)-L-arabinitol (2.00 g, 8.61 mmol). Thesuspension was heated at 50° C. for 30 minutes then cooled in anice-bath before adding the propargyl bromide (2.56 g, 17.2 mmol). Afterstirring at 0° C. for 30 mins, the reaction mixture was warmed to roomtemp and stirred for another hour before working up by carefully addingwater then evaporating the organic solvents away. The residue wasdissolved in a mixture of DCM and aqueous citric acid, took organiclayer and dried over sodium sulfate and evaporated to give an orange oil2.38 g. Silica gel chromatography eluting with DCM to 10% diethyl etherin DCM gradient gave 1.47 g of a yellow oil. This material was subjectedto another silica gel column eluting with 15% EtOAc in Petrol to give onevaporation2,3:4,5-bis-O-(1-methylethylidene)-1-O-2-propynyl-L-arabinitol (1.26 g,54%) as a colourless oil.

¹H NMR (400 MHz, CDCl₃) δ m. 4.23 (2H, OCH₂CCH), m. 4.15-4.02 (3H), dd.3.95 (1H), dd. 3.82 (1H), t. 3.71 (1H), dd. 3.63 (1H), t. 2.42 (1H,OCH₂CCH), s. 1.405 (3H), s. 1.40 (3H), s. 1.37 (3H), s. 1.33 (3H).

¹³C NMR (400 MHz, CDCl₃) δ 109.97, 109.78, 79.60, 79.60, 77.84, 77.24,74.76, 70.43, 67.77, 58.81, 27.16, 27.12, 26.85, 25.36.

1-Opropynyl-L-arabinitol (Compound 249)

2,3:4,5-bis-O-(1-methylethylidene)-1-O-2-propynyl-L-arabinitol (1.26 g,4.66 mmol) was dissolved in a mixture of TFA (4 mL) and water (2 mL).After 3 hours the solvent was evaporated and the residue dissolved inmethanol with heating. After evaporating and redissolving several timesin methanol, the residue was dissolved in the minimum amount of hotmethanol and allowed to crystallise. The crystals were filtered andwashed with a little cold methanol to give 1—O—2—propynyl—L—arabinitol(135 mg, 15%) as fine white crystals.

¹H NMR (400 MHz, D₂O) δ m. 4.22 (2H, OCH₂CCH), m. 4.03 (1H), dd. 3.79(1H), m. 3.73-3.59 (4H), m. 3.52 (1H), t. 2.86 (1H, OCH₂CCH).

¹³C NMR (400 MHz, D₂O) δ 79.55, 76.07, 71.58, 70.97, 70.87, 68.51,63.03, 58.24.

Compound 250

Compound 233 (25 mg, 0.015 mmol), sodium ascorbate (11.6 mg, 0.058 mmol)and Compound 249 (16.7 mg, 0.088 mmol) were dissolved in degassed THF (5ml) and water (2 mL). To this was added a solution of copper sulfate(10.9 mg, 0.044 mmol) in water (0.5 mL), whereupon the blue coppercolour rapidly turned to brown then faded over a few seconds to give acolourless solution. After 1 minute the solution started to turn cloudythen over an hour an orange solid had formed. The reaction mixture wasevaporated to dryness and triturated in a mixture of DCM and methanolbefore loading the supernatant onto a normal phase column eluting withan increasing gradient (0 to 50%) methanol in DCM, however thesolubility of the compound gave poor recovery of impure material. Thisimpure material was purified by reverse phase chromatography elutingwith a water-methanol gradient, and freeze-dried to give Compound 250 (7mg, 20%) as a white solid.

HRMS: (nanospray⁺) calculated for C₁₀₅H₁₅₈N₂₄O₃₃ ²⁺ [M+2H]²⁺ 1142.0726,found: 1142.0708.

Monocyclic Receptor Synthesis Scheme 2 — Synthetic Procedure Used toPrepare the Anthracene Diamine 37.

Tetra-hydroxy anthracene (33) was prepared according to literatureprocedure as described in J. Org. Chem., 1989, 54, 1018.

Tetra-tert-butyl-2,2′,2″,2‴-((9,10-dimethylanthracene-2,3,6,7-tetrayl)tetrakis(oxy))tetraacetate(34)

Under an inert N₂ amosphere, tetra-hydroxy anthracene 33 (2.35 g, 8.7mmol) was dissolved in anhydrous THF (500 mL). K₂CO₃ (4.9 g, 35.2 mmol)and tert-butyl bromoacetate (7 mL, 47.4 mmol) were added and thereaction mixture stirred under reflux for 16 hours. The mixture wascooled to room temperature and the solvent removed under vacuum. Thecrude residue was then dissolved in CH₂Cl₂ (500 mL) and washed withwater (150 mL), brine (200 mL) and dried (MgSO₄). The solvent wasremoved under vacuum and the crude residue purified by flash columnchromatography (1% MeOH:CH₂Cl₂) to yield 34 (3.8 g, 5.2 mmol, 60%) as ayellow solid. ¹H NMR: (400 MHz, (CDCl₃): δ 1.49 (s, 36H, 3 × C(1)H₃),2.85 (s, 6H, 2 x C(9)H₃), 4.76 (s, 8H, C(4)H₂), 7.38 (s, 4H, 4 × C(6)H);¹³C NMR: (100 MHz, (CDCl₃): δ 14.6 (C(9)H₃), 28.1 (C(1)H₃), 66.6(C(4)H₂), 82.3 (C(2)(CH₃)₃), 105.7 (C(6)H), 124.3 (C8), 126.2 (C7),147.2 (C5), 167.8 (C(3)O); V _(max) 2987, 2901, 1750, 1453, 1369, 1145,1066 cm⁻¹; HRMS: (ESI⁺) Found [M+Na]⁺: 749.3520.

Tetra-tert-butyl-2,2′,2″,2‴-((9,10-bis(bromomethyl)anthracene-2,3,6,7-tetrayl)tetrakis(oxy))tetraacetate (35)

Under an inert N₂ atmosphere, 34 (3 g, 4.1 mmol) was dissolved inanhydrous CH₂Cl₂ (500 mL). NBS (1.84 g, 10.3 mmol) and ABCN (50 mg, 5mol%) were added, and the mixture stirred under reflux for 1.5 hours.The reaction mixture was then cooled to room temperature and dilutedwith CH₂Cl₂ (300 mL). The solution was washed with NaOH (300 mL, 1 M),water (300 mL) and the solvent removed under vacuum to yield 35 (3.5 g,4.0 mmol, 98%) as an orange solid. ¹H NMR: (400 MHz, (CDCl₃): δ 1.52 (s,36H, 3 x C(1)H₃), 4.81 (s, 8H, C(4)H₂), 5.22 (s, 4H, 2 x C(9)H₂), 7.38(s, 4H, 4 × C(6)H); ¹³C NMR: (100 MHz, (CDCl₃): δ 28.1 (C(1)H₃), 29.7(C(9)H₃), 66.4 (C(4)H₂), 82.6 (C(2)(CH₃)₃), 104.2 (C(6)H), 126.2 (C8),126.4 (C7), 148.7 (C5), 167.4 (C(3)O); V _(max) 2987, 2933, 1706, 1488,1362, 1228, 1183, 1066 cm⁻¹; HRMS: (ESI⁺) Found [M+Na]⁺: 905.1709,907.1692.

Tetra-tert-butyl-2,2′,2”,2‴-((9,10-bis(azidomethyl)anthracene-2,3,6,7-tetrayl)tetrakis(oxy))tetraacetate (36)

Under an inert N₂ atmosphere, 35 (3.5 g, 4.0 mmol) was dissolved inanhydrous MeCN (300 mL). NaN₃ (1 g, 15.9 mmol) was added and thereaction stirred under reflux for 3 hours. The reaction mixture wascooled to room temperature and the solvent removed under vacuum. Thecrude product was dissolved in CH₂Cl₂ (400 mL), washed with water (3 ×100 mL) and the solvent removed under vacuum to yield 36 (3.2 g, 3.9mmol, 98%) as an orange solid. ¹H NMR: (400 MHz, (CDCl₃): δ 1.52 (s,36H, 3 x C(1)H₃), 4.78 (s, 8H, C(4)H₂), 5.08 (s, 4H, 2 x C(9)H₂), 7.40(s, 4H, 4 × C(6)H); ¹³C NMR: (100 MHz, (CDCl₃): δ 28.1 (C(1)H₃), 46.9(C(9)H₃), 66.4 (C(4)H₂), 82.6 (C(2)(CH₃)₃), 104.6 (C(6)H), 124.2 (C8),126.8 (C7), 148.7 (C5), 167.5 (G(3)O); v max 2988, 2931, 2091, 1736,1498, 1364, 1227, 1186, 1062 cm-¹; HRMS: (ESI⁺) Found [M+Na]⁺: 831.3530.

Tetra-tert-butyl-2,2′,2“,2‴-((9,10-bis(aminomethyl)anthracene-2,3,6,7-tetrayl)tetrakis(oxy))tetraacetate (37)

Under an inert N₂ atmosphere, 36 (100 mg, 0.12 mmol) was dissolved inanhydrous degassed THF (8 mL). PMe₃ was added (2.5 mL, 2.5 mmol, 1 M inTHF) and the mixture stirred at room temperature for 3 hours. Degassedwater (2 mL) was added and the reaction mixture stirred for 1 hour. Thesolvent was then evaporated under a flow of nitrogen and the cruderesidue dissolved in THF/H₂O (5:1, 3 mL). The solvent was then removedby freeze-drying to yield 37 (90 mg, 0.12 mmol, 96%) as a pale brownsolid. ¹H NMR: (400 MHz, (CDCl₃): δ 1.50 (s, 36H, 3 × C(1)H₃), 4.58 (s,4H, 2 x C(9)H₂), 4.77 (s, 8H, C(4)H₂), 7.49 (s, 4H, 4 x C(6)H); ¹³C NMR:(100 MHz, (CDCl₃): δ 28.1 (C(1)H₃), 38.9 (C(9)H₂), 66.5 (C(4)H₂), 82.4(C(2)(CH₃)₃), 105.0 (C(6)H), 125.8 (C8), 126.2 (C7), 148.0 (C5), 167.6(G(3)O); ); v max 2982, 2926, 1729, 1497, 1358, 1222, 1144, 1069 cm⁻¹;HRMS: (MALDI⁺) Found [M+H]⁺: 757.3909.

Scheme 3 — Synthetic Procedure Used to Prepare the Monocylic Receptor40.

Tetra-tert-butyl-2,2′,2“,2‴-((9,10-bis(isocyanatomethyl)anthracene-2,3,6,7-tetrayl)tetrakis(oxy))tetraacetate (38)

Under an inert N₂ atmosphere, a flask was charged with 37 (30 mg, 0.04mmol) and NaHCO₃ (12 mg, 0.14 mmol). CH₂Cl₂ (1 mL) and H₂O (1 mL) wereadded, the mixture cooled to 0° C. and rapidly stirred. Triphosgene (9.4mg, 0.016 mmol) was added and the reaction mixture stirred at roomtemperature for 30 minutes. The reaction mixture was diluted with CH₂Cl₂(10 mL) and the organic layer separated, dried (MgSO₄) and the solventremoved under vacuum to afford 38 (29 mg, 0.036 mmol, 91%) as an orangesolid. ¹H NMR: (400 MHz, (CDCl₃): δ 1.51 (s, 36H, 3 × C(1)H₃), ), 4.78(s, 8H, C(4)H₂), 5.07 (s, 4H, 2 x C(9)H₂), 7.35 (s, 4H, 4 × C(6)H); ¹³CNMR_(:) (100 MHz, (CDCl₃): δ 28.0 (C(1)H₃), 39.83 (C(9)H₂), 66.6(C(4)H₂), 82.7 (C(2)(CH₃)₃), 104.4 (C(6)H), 125.5 (C8), 126.0 (C7),148.7 (C5), 167.3 (G(3)O); V _(max) 2979, 2934, 2251, 1734, 1493, 1367,1225, 1144, 1064 cm-¹; HRMS: (ESI⁺) Found [M+Na]⁺: 831.3319.

Tert-butyl Protected Half Receptor (39)

Under an inert N₂ atmosphere, 1,2-phenylene diamine (0.3 g, 2.70 mmol)was dissolved in dry degassed CH₂Cl₂ (120 mL). A solution of 38 (55 mg,0.068 mmol) in dry degassed CH₂Cl₂ (50 mL) was added dropwise over 10minutes and then stirred at room temperature for 30 minutes. The solventwas removed under vacuum and the crude solid purified by flash columnchromatography (80:20 EtOAc:hexane ➔ 5:95 MeOH:CH₂CI₂ ➔ 10:90MeOH:CH₂Cl₂) to yield 39 (53 mg, 0.052 mmol, 77%) as an orange brownsolid. ¹H NMR: (400 MHz, ((CD₃)₂SO): δ 1.47 (s, 36H, 3 × C(1)H₃), ),4.59 (s, 4H, N(19)H₂), 4.87 (s, 8H, C(4)H₂), 5.12 (s, 4H, 2 x C(9)H₂),6.50-6.57 (m, 4H, C(13)H and N(18)H), 6.66-6.70 (m, 2H, C(15)H), 6.77(t, J = 7.6 Hz, 2H, C(14)H), 7.42-7.48 (m, 4H, C(12)H and N(18)H), 7.66(s, 4H, 4 × C(6)H); ¹³C NMR: (100 MHz, ((CD₃)₂SO): δ 28.2 (C(1)H₃),36.7(C(9)H₂), 66.8 (C(4)H₂), 82.9 (C(2)(CH₃)₃), 105.2 (C(6)H), 114.5(C15), 118.9 (C13), 122.8 (C11), 125.2, 125.5 (C12 and C14), 125.8 (C8),126.2 (C7), 148.3 (C5), 149.5 (C16), 154.2 (C10), 167.5 (G(3)O); V_(max) 3315, 2973, 2901, 1733, 1622, 1494, 1393, 1225, 1146, 1057, 742cm-¹; HRMS: (ESI⁺) Found [M+Na]⁺: 1047.4689.

Tert-butyl Protected Tetra-urea Macrocycle (40)

Under an inert N₂ atmosphere, 38 (40 mg, 0.049 mmol) was dissolved indry degassed CH₂Cl₂ (600 mL). To this was added a solution of 39 (50 mg,0.049 mmol) in dry degassed pyridine (60 mL) dropwise over 20 minutes.The reaction was stirred at room temperature for 16 hours and then thesolvent removed under reduced pressure. The crude residue was suspendedin HPLC grade water and freeze-dryed to afford a fine crude solid. Theproduct was then purified by reverse phase HPLC and freeze dried toafford 40 (50 mg, 0.027 mmol, 56%) as an off white solid. ¹H NMR: (400MHz, ((CD₃)₂CO): δ 1.49 (s, 36H, 3 x C(1)H₃), ), 4.78 (m, 16H, C(4)H₂),5.12 (m, 8H, C(9)H₂), 6.95 (s, 4H, C(13)H), 7.65 (s, 8H, C(6)H), 8.07(s, 4H, C(12)H); ¹³C NMR: (100 MHz, ((CD₃)₂CO): δ 27.3 (C(1)H₃), 36.0(C(9)H₂), 66.0 (C(4)H₂), 81.6 (C(2)(CH₃)₃), 105.3 (C(6)H), 126.3 (C8),126.9 (C7), 127.4 (C12), 132.2 (C13), 135.6 (C11), 147.8 (C5), 155.5(C10), 167.9 (G(3)O); HRMS: (ESI⁺) Found [M+Na]⁺: 1856.8145, [M+2Na]²⁺:939.9019.

Scheme 4 — Synthetic Procedure Used to Prepare the Monocylic Receptor89.

9,10-Bis(isocyanatomethyl)anthracene (55)

Under an inert N₂ atmosphere, a flask was charged withanthracene-9,10-diyldimethanamine An-NH₂* (20 mg, 0.085 mmol) and NaHCO₃(26 mg, excess). CH₂Cl₂ (1 mL) and H₂O (1 mL) were added, the mixturecooled to 0° C. and rapidly stirred. Triphosgene (20 mg, 0.068 mmol) wasadded and the reaction mixture stirred at room temperature for 30minutes. The reaction mixture was diluted with CH₂Cl₂ (10 mL) and theorganic layer separated, dried (MgSO₄) and the solvent removed undervacuum to afford 55 (22 mg, 0.076 mmol, 91%) as a yellow solid. ¹H NMR:(400 MHz, (CDCl₃): δ 5.38 (s, 4H, C(5)H₂), 7.66 (dd, J = 6.9, 3.2 Hz,4H, C(1)H), 8.35 (dd, J = 6.9, 3.2 Hz, 4H, C(2)H); ¹³C NMR: (100 MHz,(CDCl₃): δ 39.0 (C(5)H₂), 124.1 (C(1)H), 126.7 (G(8)H), 127.3 (C4),129.3 (C3), 167.3; V _(max) 2921, 2234, 1620, 1491, 1448, 1324, 1185,858, 751 cm-¹; HRMS: (ESI⁺) Found [M+Na]⁺: 311.0785.

* prepared according to the synthetic procedures described in literatureprocedure as described in Org. Biomol. Chem., 2005, 3, 48.

Diamino Tert-butyl Protected Anthracene Half Receptor (88)

Under an inert N₂ atmosphere, 84 (350 mg, 0.195 mmol) was dissolved inanhydrous dichloromethane (10 mL). 55 (25 mg, 0.098 mmol) was added andthe reaction heated to reflux for 2 days. The reaction was cooled toroom temperature and the solvent removed under vacuum. The crude residuewas purified by reverse phase HPLC to afford the Fmoc protected product86 (254 mg, 0.66 mmol, 67%) as a white solid. Conversion to 86 wasconfirmed by limited NMR studies* and high resolution mass spectrometry(ESI⁺): m/z calculated for [M+2Na]²⁺ 1963.1077, found 1963.1067. Underan inert N₂ atmosphere, 86 was dissolved in anhydrous dichloromethane(10 mL) and cooled to 0° C. DBU (45 µL, 0.28 mmol) was added dropwiseand the reaction mixture warmed to room temperature and stirred for 2hours. The solvent was removed under vacuum and the crude productpurified by flash column chromatography (5% MeOH:CH₂Cl₂) to afford 88(215 mg, 0.063 mmol, 95%) as an off-white solid. ¹H NMR: (400 MHz,(CD₃OD): δ 1.43 (s, 162H, C(23)H₃), 1.93 (m, 36H, C(20)H₂), 2.08 (m,12H, C(15)H₂), 2.18 (m, 48H, C(19, 16)H₂), 5.36 (s, 4H, C(5)H₂), 7.18(dd, J = 2.1, 8.3 Hz, 2H, C(9)H), 7.26 (d, J = 2.1 Hz, 2H, C(11)H), 7.41(d, J = 8.3 Hz, 2H, C(8)H), 7.41 (s, 6H, NH), 7.60 (dd, J = 3.3, 6.9 Hz,4H, C(1)H), 7.89 (s, 2H, NH), 8.47 (dd, J = 3.3, 6.9 Hz, 4H, C(2)H); ¹³CNMR: (100 MHz, (CDCI₃): δ 27.1 (C23), 29.1 (C20), 29.3 (C21), 31.0(C15), 31.1 (C16), 57.4 (C18), 58.0 (C14), 80.2 (C22), 115.9 (C11),117.4 (C9) 123.1 (C8), 124.6 (C2), 125.9 (C1), 128.5 (C10), 130.1 (C4),130.5 (C3), 131.4 (C7), 140.0 (C12), 156.7 (C6), 168.6 (C13), 173.0(C21), 174.1 (C17); HRMS: (ESI⁺) Found [M+H+Na]²⁺: 1730.5507.

* Limited NMR studies were only possible due to believed slowconformational exchange of 86 resulting in very broad signals of lowintensity.

Tert-butyl Protected G2 Anthracene Tetra Urea Macrocycle (89)

Under an inert N₂ atmosphere, 55 (5.3 mg, 0.018 mmol) was dissolved inanhydrous degassed dichloromethane (600 mL) and heated to reflux. 88 (63mg, 0.018 mmol) in anhydrous degassed dichloromethane (50 mL) was addedover 30 mins and stirred at reflux for 4 days. The solvent was thenremoved under vacuum and the crude product purified by reverse phaseHPLC and then freeze dried to afford 89 (25 mg, 6.7 µmol, 37%) as awhite solid. ¹H NMR: (400 MHz, (CD₃OD): δ 1.45 (s, 162H, C(23)H₃), 1.99(m, 36H, C(20)H₂), 2.17 (m, 12H, C(15)H₂), 2.24 (m, 48H, C(19)H₂), 2.31(m, 12H, C(16)H₂), 5.39 (s, 8H, C(5)H₂), 7.31, 7.44 (br s, 4H, C(1)H),7.49 (s, 6H, NH), 7.70 (dd, J = 2.1, 8.5 Hz, 2H, C(9)H), 7.89 (d, J =8.5 Hz, 2H, C(8)H), 7.98 (s, 2H, C(11)H), 8.39 (br s, 8H, C(2)H); ¹³CNMR: (100 MHz, (CDCl₃): δ 27.0 (C23), 29.1 (C20), 29.3 (C21), 30.8(C15), 31.1 (C16), 57.3 (C18), 58.1 (C14), 80.3 (C22), 121.6 (C9),121.77 (C12) 124.4 (C2), 124.7 (C11), 125.8, 125.9 (C1), 130.0 (C10),130.4 (C3), 130.6 (C3), 131.4 (C7, 12), 156.0, 156.8 (C6), 168.1 (C13),173.1 (C21), 174.1, 174.2 (C17); HRMS: (ESI⁺) Found [M+3Na]³⁺:1264.3835, [M+4Na]⁴⁺: 954.0378.

Fmoc Protected Methyl Ester G2 Linker (94)

84 (215 mg, 0.12 mmol) was dissolved in dichloromethane (5 mL) and TFA(5 mL) added dropwise. The solution was stirred at room temperature for16 hours and the TFA and solvent evaporated under a flow of N₂. Theresidue was redissolved in methanol (5 mL) and trimethyl orthoformate (5mL). HCl (5% v/v, 0.5 mL) was added and the mixture stirred for 24hours. The solvent was then removed under vacuum and the crude productpurified by flash column chromatography (5% MeOH:CH₂Cl₂) to afford 94(147 mg, 0.10 mmol, 87%). ¹H NMR: (400 MHz, (CDCl₃): δ 1.99 (m, 18H,C(23)H₂), 2.108(m, 6H, C(18)H₂), 2.25 (m, 24H, C(22, 19)H₂), 3.60 (s,27H, C(25)H₃), 4.23 (m, 1H, C(7)H), 4.48 (m, 2H, C(8)H₂), 6.31 (s, 3H,NH), 6.72 (d, J = 8.4 Hz, 1H, C(13)H), 7.26 (m, 3H, C(4, 12)H), 7.38 (t,J = 7.4 Hz, 2H, C(3)H), 7.63 (m, 2H, C(5)H), 7.75 (d, J = 7.9 Hz, 3H,C(2, 15)H), 8.12 (s, 1H, NH); ¹³C NMR: (100 MHz, (CDCl₃): 28.3 (C22),29.6 (C23), 31.8 (C19), 32.2 (C18), 47.2 (C7), 51.8 (C25), 57.3 (C21),58.2 (C17), 67.1 (C8), 116.4 (C12), 120.0 (C2), 122.6 (C10), 124.4(C14), 125.1 (C4), 125.2 (C15), 126.6 (C13), 127.7 (C5), 127.8 (C3),141.3 (C1), 143.6 (C11), 143.8 (C6), 155.0 (C9), 166.6 (C16), 173.3(C20), 173.8 (C24); V max 3330, 2976, 2961, 1727, 1658, 1531, 1452,1367, 1249, 1150, 846 cm⁻¹; HRMS: (ESI⁺) Found [M+2Na]²⁺: 731.3279,[M+Na]⁺: 1439.6353.

Diamino Methyl Ester Protected Anthracene Half Receptor (97)

Under an inert N₂ atmosphere, 94 (160 mg, 0.11 mmol) was dissolved inanhydrous dichloromethane (5 mL). 55 (15 mg, 0.054 mmol) was added andthe reaction heated to reflux for 4 days. The reaction was cooled toroom temperature and the solvent removed under vacuum. The crude residuewas purified by flash column chromatography (5% MeOH:CH₂Cl₂) to affordthe Fmoc protected product 97a (190 mg, 0.061 mmol, 56%) as a whitesolid. Conversion to 97a was confirmed by limited NMR studies* and highresolution mass spectrometry (ESI⁺): m/z calculated for [M+2Na]²⁺1584.1834, found 1584.1849. Under an inert N₂ atmosphere, 97a wasdissolved in anhydrous dichloromethane (10 mL) and cooled to 0° C. DBU(50 µL, 0.30 mmol) was added dropwise and the reaction mixture warmed toroom temperature and stirred for 4 hours. The solvent was removed undervacuum and the crude product purified by flash column chromatography (4%MeOH:CH₂Cl₂) to afford 97 (151 mg, 0.057 mmol, 93%) as an off whitesolid. ¹H NMR: (400 MHz, (CD₃OD): δ 1.94 (m, 36H, C(20)H₂), 2.07 (m,12H, C(15)H₂), 2.24 (m, 48H, C(19, 16)H₂), 3.60 (s, 54H, C(22)H₃), 4.94(s, 4H, C(5)H₂), 7.05 (d, J = 8.3 Hz, 2H, C(9)H), 7.15 (m, 4H, C(11,8)H), 7.37 (s, 6H, NH), 7.48 (dd, J = 3.3, 6.9 Hz, 4H, C(1)H), 7.85 (s,2H, NH), 8.21 (dd, J = 3.3, 6.9 Hz, 4H, C(2)H); ¹³C NMR: (100 MHz,(CDCl₃): δ 28.5 (C20), 29.3 (C21), 31.0 (C15), 31.1 (C16), 51.8 (C22),57.3 (C18), 58.1 (C14), 116.1(C11), 117.5 (C9) 123.0 (C8), 124.4 (C2),125.8 (C1), 128.6 (C10), 130.0 (C4), 130.5 (C3), 131.4 (C7), 140.1(C12), 155.5 (C6), 166.7 (C13), 173.3 (C17), 173.9 (C21); HRMS: (ESI⁺)Found [M+2Na]²⁺: 1361.1096, Found [M+3Na]³⁺: 915.7415.

* Limited NMR studies were only possible due to believed slowconformational exchange of 97a resulting in very broad signals of lowintensity.

Methyl Ester Protected G2 Anthracene Tetra Urea Macrocycle (98)

Under an inert N₂ atmosphere, 55 (7.9 mg, 0.027 mmol) was dissolved inanhydrous degassed dichloromethane (600 mL) and heated to reflux. 97 (73mg, 0.027 mmol) in anhydrous degassed dichloromethane (50 mL) was addedover 30 mins and stirred at reflux for 5 days. The solvent was thenremoved under vacuum and the crude product purified by reverse phaseHPLC and then freeze dried to afford 98 (21 mg, 7.0 µmol, 26%) as a paleyellow solid. ¹H NMR: (400 MHz, (CD₃)₂SO): δ 1.89 (m, 36H, C(20)H₂),1.95 (m, 12H, C(15)H₂), ), 2.12 (m, 12H, C(16)H₂), 2.22 (m, 36H,C(19)H₂), 5.25 (s, 8H, C(5)H₂), 7.32 (s, 6H, NH), 7.42 (br s, 4H,C(1)H), 7.52 (m, 6H, C(1, 9)H), 7.74 (s, 2H, NH), 7.87 (d, J = 8.5 Hz,2H, C(8)H), 8.07 (s, 2H, C(11)H), 8.37 (br s, 8H, C(2)H); ¹³C NMR: (100MHz, (CD₃)₂SO): δ 28.3 (C20), 29.2 (C21), 29.5, 29.8 (C15), 30.7, 31.0(C16), 35.7 (C5), 51.8 (C22), 56.8 (C18), 57.9 (C14), 120.0 (C8), 122.8(C9), 123.3 (C11), 125.4 (C2), 125.7 (C12), 126.4, 126.5 (C1), 129.0,129.5 (C3), 129.9 (C4), 135.6 (C7) 155.1, 155.7 (C6), 166.0 (C13), 172.9(C17), 173.7 (C21); HRMS: (ESI⁺) Found [M+2Na]²⁺: 1506.1621, [M+3Na]³⁺:1011.7687.

Deprotected Anthracene Tetra Urea Macrocycle (90)

Protected receptor 89 (30 mg, 0.008 mmol) was dissolved indichloromethane (HPLC grade, 2.7 ml) and cooled to 0° C. Trifluoroaceticacid (TFA) (0.3 mL) was added dropwise and the reaction warmed to roomtemperature and stirred for 4 hours. The solvent was then removed undera flow of nitrogen, then the residue was co-evaporated with toluene (3 x10 mL) to remove residual TFA, suspended in water and freeze dried. Theproduct was then purified by preparative HPLC (Waters CSH C18 5 µm 19 ×250 mm) eluting with 100% Water (buffered with 0.1 % TFA) ➔ 100%methanol over 40 minutes. The solvent was removed under vacuum, theresidue co-evaporated with toluene (3 × 10 mL), the product suspended inwater and freeze dried. The solid was then suspended in water,neutralised to pH 7.4 with NaOH (aq), filtered and then freeze dried toafford 90 as a white solid (21 mg, 0.0068 mmol, 85%). ¹H NMR: (400 MHz,75° C., D₂O): δ 2.40-2.50 (m, 36H, C(19)H₂), 2.58-2.71 (m, 48H, C(20)H₂,C(15)H₂), 2.80-2.90 (m, 12H, C(16)H₂), 5.63, 5.81 (br s, 4H, C(5)H₂),7.43-7.50, 7.98-8.06 (br m, 4H, C(1)H),8.25 (br s, 2H, C(11)H), 8.27 (d,2H, C(9)H), 8.39 (d, 2H, C(8)H), 7.63 (d, J = 8.3 Hz, 3H, C(10)H),8.58-8.65, 8.82-8.89 (br m, 4H, C(2)H).

Scheme for Tetra-methoxy Anthracene Isocyanate (95)

2,3,6,7-Tetramethoxy-9,10-bis(bromomethyl)anthracene, (92)

Under an inert N₂ atmosphere, 91 (2 g, 6.1 mmol), NBS (4 g, 22.6 mmol)and ABCN (73 mg, 0.3 mmol) were dissolved in anhydrous dichloromethane(150 mL), and the mixture stirred at reflux for 4 hours. The mixture wasthen cooled to 0° C. and filtered. The solid was then dried under highvacuum to afford 92 (2.1 g, 4.3 mmol, 71%) as a bright yellow solid. ¹HNMR (400 MHz, CDCl₃) δ 4.11 (s, 12H, C(1)H), 5.35 (s, 4H, C(6)H), 7.40(s, 4H, C(3)H), ¹³C NMR (100 MHz, CDCl₃) 28.7 (C6), 56.0 (C1), 101.8(C3), 125.7 (C5), 125.9 (C4), 150.1 (C2).

2,3,6,7-Tetramethoxy-9,10-bis(azidoomethyl)anthracene, (93)

Under an inert N₂ atmosphere, 92 (2.7 g, 5.58 mmol) and NaN₃ weresuspended in anhydrous MeCN (70 mL). The mixture was stirred at refluxfor 16 hours and then cooled to room temperature and the solvent wasevaporated in vacuo. The remaining residue was then suspended in water(200 mL) and filtered. The solid was washed with ethanol (3 × 100 mL)and dried under high vacuum to afford 93 (1.6 g, 3.9 mmol, 70%) as abrown solid. ¹H NMR (400 MHz, CDCl₃) δ 4.10 (s, 12H, C(1)H), 5.19 (s,4H, C(6)H), 7.42 (s, 4H, C(3)H), ¹³C NMR (100 MHz, CDCl₃) 47.7 (C6),56.1 (C1), 101.8 (C3), 123.8 (C5), 126.9 (C4), 150.2 (C2); HRMS: (ESI⁺)Found [M+Na]⁺431.1448.

2,3,6,7- Tetramethoxy-9,10-bis(aminomethyl)anthracene, (94)

Under an inert N₂ atmosphere, azide 93 (1.6 g, 3.90 mmol) and PPh₃ (8 g,31.4 mmol) were suspended in degassed THF (80 mL). Degassed water (4 mL)was added and the reaction heated to 60° C. for 16 hours. The reactionwas cooled to room temperature and the solvent removed under vacuum. Thecrude residue was suspended in toluene (200 mL), filtered, washed withtoluene (2 × 100 mL) and dried under high vacuum to afford 94 (1.05 g,2.96 mmol, 76%) as a pale brown solid. ¹H NMR (400 MHz, CDCl₃) δ 4.08(s, 12H, C(1)H), 4.69 (s, 4H, C(6)H), 7.49 (s, 4H, C(3)H), ¹³C NMR (100MHz, CDCl₃) δ 39.5 (C6), 56.0 (C1), 101.9 (C3), 125.3 (C5), 125.5 (C4),149.7 (C2); HRMS: (ESI⁺) Found [M+Na]⁺ 379.1626.

9, 10-Bis(isocyanatomethyl)-2,3,6,7-tetramethoxyanthracene (95)

Under an inert N2 atmosphere, a flask was charged with triphosgene (324mg, 1.1 mmol) and anhydrous toluene (15 mL) was added A suspension of 94(200 mg, 0.55 mmol) in anhydrous toluene (5 mL) was added dropwise andthe reaction mixture stirred at reflux for 2 hours. The reaction mixturewas cooled and the solvent removed under high vacuum. The crude solidwas resuspended in dichloromethane (50 mL) and filtered. The filtratewas collected and the solvent removed under vacuum to afford 95 (121 mg,0.30 mmol, 54%) as a brown solid. ¹H NMR (400 MHz, CDCl₃) δ 4.11 (s,12H, C(1)H), 5.21 (s, 4H, C(6)H), 7.37 (s, 4H, C(3)H), V _(max) 2934,2832, 2255, 1498, 1435, 1245, 1204, 1169, 1028 cm⁻¹; HRMS: (ESI⁺) Found[M+Na]⁺ 431.1218.

Scheme for Octa-methoxy Anthracene Bis Urea Receptor

Diamino Tert-butyl Protected Methoxy-anthracene Half Receptor (96)

Under an inert N₂ atmosphere, 84 (507 mg, 0.283 mmol) and 94 (50 mg,0.13 mmol) were dissolved in anhydrous dichloromethane (8 mL). Pyridine(60 µL, 0.74 mmol) was added and the reaction heated to reflux for 16hours. The reaction was cooled to room temperature and the solventremoved under vacuum. The crude residue was purified by reverse phaseHPLC to afford the Fmoc protected product 95 (336 mg, 0.84 mmol, 65%) asa white solid. Conversion to 95 was confirmed by limited NMR studies*and high resolution mass spectrometry (ESI⁺): m/z calculated for[M+2Na]²⁺ 2015.6171, found 2015.6176. Under an inert N₂ atmosphere, 95(100 mg, 0.028 mmol) was dissolved in anhydrous dichloromethane (10 mL)and cooled to 0° C. DBU (50 µL, 0.31 mmol) was added and the reactionmixture warmed to room temperature and stirred for 1 hour. The solventwas removed under vacuum and the crude product purified by flash columnchromatography (6% MeOH:CH₂Cl₂) to afford 96 (91 mg, 0.026 mmol, 92%) asan off white solid. ¹H NMR: (400 MHz, (CD₃OD): δ 1.42 (s, 162H,C(23)H₃), 1.84-2.0 (m, 36H, C(20)H₂), 2.02-2.14 (m, 12H, C(15)H₂),2.13-2.31 (m, 48H, C(19, 16)H₂), 3.95 (br s, 12H, C(24)H₃), 4.58 (br s,4H, C(5)H₂), 7.13 (d, J = 8.3 Hz, 2H, C(9)H), 7.23 (s,2H, C(11)H), 7.31(d, J = 8.3 Hz, 2H, C(8)H), 7.38 (br s, 4H, C(2)H), 7.41 (s, 6H, NH),7.89 (s, 2H, NH); ¹³C NMR: (100 MHz, (CDCl₃): δ 27.1 (C23), 29.1 (C20),29.3 (C21), 30.8 (C15), 31.1 (C16), 54.9 (C24), 57.4 (C18), 58.1 (C14),80.3 (C22), 101.9 (C2), 115.8 (C11), 117.3 (C9) 123.3 (C8), 126.1 (C4),126.1 (C3), 128.2 (C10), 131.6 (C7), 140.3 (C12), 149.5 (C1), 157.0(C6), 168.5 (C13), 173.0 (C21), 174.1 (C17); HRMS: (ESI⁺) Found[M+2Na]²⁺:1208.3682.

* Limited NMR studies were only possible due to believed slowconformational exchange of 95 resulting in very broad signals of lowintensity.

Tert-butyl Protected Octa-methoxy Anthracene Tetra Urea Macrocycle (97)

Under an inert N₂ atmosphere, 96 (25 mg, 0.007 mmol) and DMAP (1.7 mg,0.014 mmol) were dissolved in anhydrous degassed dichloromethane (12 mL)and heated to reflux. 94 (2.7 mg, 0.007 mmol) in anhydrous degasseddichloromethane (2 mL) was added and the reaction stirred at reflux for2 days. The solvent was then removed under vacuum and the crude productpurified by reverse phase HPLC and then freeze dried to afford 89 as awhite solid.

Binding Studies

Isothermal titration calorimetry (ITC) and ¹H NMR were used to determinethe binding affinities between the compounds of the present invention(e.g. receptor compound 1 and receptor compound 90) and a number ofsaccharides (e.g. glucose, mannose and cellobiose), together with othersmall molecules (e.g. uracil and uric acid). ITC and ¹H NMR titrationswere performed according to the general procedure described hereinaboveand the ITC traces, ¹H NMR spectra and binding affinities are summarisedin FIGS. 2 to 75 .

¹H NMR Titrations

¹H-NMR titrations were performed on a Varian VNMR cryogenically cooledS600 spectrometer. Solutions of saccharides in D₂O (99.9%), containingreceptor at a known concentration to be used in the experiment, wereprepared and allowed to equilibrate overnight before use if necessary.Aliquots were then added to an NMR tube containing a known concentrationof receptor solution (typically 50 µM - 250 µM). The receptorconcentration was therefore held constant while the carbohydrateconcentration was increased. The sample tube was shaken after eachaddition, centrifuged and ¹H-NMR spectra were acquired at 298 K.

If the receptor bound saccharide slower than the NMR sample rate (“slowexchange”), the K_(a) was determined by analysing the NMR integral of apeak assigned to the Host-Guest complex. The variable X was defined asthe integral of an isolated resonance of the complex (typically in thearomatic region) divided by the integral of all the related resonances(typically the whole aromatic region). As X is proportional to fractionof host in the bound state, the change in X could be plotted as afunction of the guest concentration to give a curve which could befitted to a 1:1 binding model to yield the association constant K_(a).Mathematically, the fitting process is essentially identical to thatemployed for binding with fast exchange, except that the integral of apeak due to the complex replaces the chemical shift of a peak due tobound + unbound receptor. The calculation was performed using anon-linear least squares curve-fitting programme implemented withinExcel. The programme yields binding constants K_(a) and limiting X(X_(lim)) as output. K_(a) values are listed in Table 4 below. Anestimated error for Ka was obtained from individual data points byassuming the determined K_(a) and X_(lim). These errors are reported inTable 4 and are typically well below 5%.

TABLE 1 Relative integrations of α—H1 and β—H2 during NMR titration.Glucose concentration (µM) Relative integration (αH1:βH2) β-D-glucose(%) 57 -^(a) - 114 -^(a) - 170 1:2.08 67% 225 1:2.01 66% 280 1:1.85 64%334 1:2.04 67% 387 1:1.88 65% 440 1:1.84 64% 492 1:1.82 64% 544 1:1.8264% ^(a) values for integration not obtained due to low intensity of andbroadness of signals.

TABLE 2 Relative integrations of α—H1 and β—H2 over time from pureα-D-glucose, with and without Receptor 1 (0.2 mM) present. Time (min)Relative integration (αH1:βH2) D-glucose (5 mM) only D-glucose (5 mM)and receptor X (0.2 mM) 0 1:0.01 1:0.06 10 1:0.06 1:0.11 30 1:0.191:0.22 60 - 1:0.30 70 1:0.34 - 90 - 1:0.38 100 1:0.50 - 120 1:0.591:0.52 150 1:0.70 1:0.66 180 1:0.86 - 210 1:0.93 -

TABLE 3 Calculated values for K_(a) when titrating cellobiose (250 mM)against receptor 1 (0.11 mM). Volume of Guest added (µL) [Host]_(free) /µM [Guest]_(free) / mM [Host]_(total) / µM [Guest]_(total) / mM [Host-Guest] / µM Integral (peak 8.02 ppm •) Integral of peak • vs integral ofpeak • when [H]_(free) = 0. K_(a) / M⁻¹ 0 110 0 110 0 - 0 0 4 110 20 11020 0 0.00005 0.0011 0.55 4 99 39 110 39 11 0.00465 0.102 29.0 4 93 58110 58 17 0.007 0.154 31.2 8 86 96 110 96 24 0.01 0.220 29.4 8 75 132110 132 35 0.0143 0.315 34.9 8 47 167 110 167 63 0.0262 0.577 81.5 20 38251 110 251 72 0.0296 0.650 74.0 20 8 329 110 329 102 0.0421 0.927 38640 0 470 110 470 110 0.0455 1 - Average K_(a) / M⁻¹ 31.1 Std Dev (Error)2.66 (9%)

The integrals of the peak at 8.02 ppm (denoted with •, see FIG. 31 )were made relative to integral of the same peak (8.02 ppm) when allreceptor is assumed to be saturated with guest (i.e. the final additionin the titration, row denoted with yellow). These relative integrals arethen used to determine the amount of Host-Guest [HG]. This value for[HG] along with calculated values for free host [H]_(free) and freeguest [G]_(free) can be used to calculate the K_(a) at each point in thetitration. An average of the values obtained (denoted in blue) was thenused as the overall K_(a) (31.1 M⁻¹) along with the associated standarddeviation and error. Not all K_(a) values calculated were included inthe averaged K_(a) value. The earlier integrations are unreliable due tothe very small intensity of the peak at 8.02 ppm. The later integrationswere also deemed unreliable due to large deviations in baseline of thespectra due to the large excess of guest present. The selected valuesfor K_(a) used for the average calculation and the averaged K_(a) itselfcorroborate with the results obtained from ITC.

Isothermal Titration Microcalorimetry (ITC) Titrations

Isothermal Titration MicroCalorimetry (ITC) experiments were performedon a MicroCal iTC200 microcalorimeter and/or a MicroCali VP-ITC. ITCexperiments were carried out at 298 K. Saccharide solutions wereprepared in HPLC-grade water with 10 mM phosphate buffer solution (pH7.4) and allowed to equilibrate overnight if necessary. The sample cellwas charged with a known concentration of receptor solution inHPLC-grade water with 10 mM phosphate buffer solution at pH 7.4(typically 50 µM - 200 µM). Then, aliquots (typically 1.0 µL) ofcarbohydrate solution were added and the evolution of heat was followedas a function of time. Heats of dilution were measured by injecting thesame carbohydrate solution into HPLC-grade water with 10 mM phosphatebuffer solution at pH 7.4, using identical conditions. For everyaddition, the heat of dilution was subtracted from the heat of bindingusing a MicroCal software programme implemented in ORIGIN 7.0. This gavean XY matrix of heat vs. total guest concentration. This matrix was thenimported into a specially written Excel programme to fit the data to a1:1 binding model to give a K_(a). ΔG can be derived from K_(a) and thusΔS can be derived from ΔH and ΔG using common thermodynamic equations.The fitting procedure also yields errors in K_(a) as in the case of NMRdescribed above. This method consistently produced more accurate fitsthan fitting the data to an S-curve, as in the MicroCal software(S-curves are typically not observed for binding constants below ~104 -105 M⁻¹). Although fits produced using the supplied MicroCal softwarewere generally similar to those calculated using the Excel programme,they also consistently overestimated the K_(a) by approximately 10%. Itwas found that better corroboration of the ITC data with the equivalentNMR data was achieved using the Excel programme. ITC outputs for heat ofdilution of substrates, binding events between substrates and receptor1, and analysis curves are included in the Figures. An overview of thebinding data, including thermodynamic quantities and errors is given inTable 4 below.

TABLE 4 binding affinities of various substrates for receptor 1.Substrate (medium) Determined by NMR K_(a) (M⁻¹) Determined by ITC (kJmol⁻¹) K_(a) (M⁻¹) ΔG ΔH TΔS D—Glucose 18,026 ± 1.04% 18,600 ± 14.3%-24.4 -7.8 16.6 D—Glucose pH 6 (PBS) - 17,300 ± 3.8% -24.2 -2.6 21.6D—Glucose pH 7 (PBS) - 17,800 ± 5.5% -24.3 -2.2 22.0 D—Glucose pH 8(PBS) - 18,300 ± 1.8% -24.3 -2.6 21.8 D—Glucose (human serum) - 2477 ±5.7% -19.4 -4.1 15.3 D—Glucose (DMEM cell culture) - 5637 ± 2.1% -21.4-5.2 16.2 D—Glucose (DMEM salt control) - 5164 ± 5.9% -21.2 -5.0 16.2D—Glucose (Leibovitz’s L-15 cell culture) - 5214 ± 8.6% -21.2 -4.2 17.0Methyl ß—D—Glucoside 7522 ± 5.5% 7886 ± 16.4% -22.2 -3.2 21.2Myo—inositol 7328 ± 7.4% 7563 ± 4.2 % -22.1 -22.1 -2.4 D—Glucuronic Acidn.d.^(a) 5348 ± 3.5% -21.3 -27.8 -6.5 D—Xylose n.d.^(a) 5804 ± 3% -21.5-8.0 13.5 2—Deoxy—D—Glucose n.d.^(a) 725 ± 5.7% -16.3 -2.9 13.4D—Galactose 132 ± 10% 182 ± 2.3% -12.9 -8.8 4.2 D—Mannose 140 ± 1.3% 143± 1.1% -12.3 -11.8 0.6 D—Ribose 267 ± 3.8% 216 ± 1.9% -13.3 -23.0 -9.7D—Fructose 51 ± 5.5% 60 ± 2.7% -10.6 -20.0 -9.5 D—Cellobiose 31 ± 9% 30± 15.9% -8.5 -9.2 -0.7 Mannitol - 0 Gluconate^(b) 0 0 Methylα—D—Glucoside 0 0 N—Acetyl—D—glucosamine - 0 D—Maltose - 0 L—Fucose - 0Uracil (PBS) - 0 Uric Acid (PBS) - 0 Cytosine - 0 Adenosine - 0Paracetamol - 0 Ascorbic Acid - 0 L—Phenylalanine - 0 L—Tryptophan - 0All solutions at pH 7.4 in 10 mM Phosphate buffer solution unlessotherwise stated. Human blood Serum and cell culture media were dialysedat 10k MWCO and then buffered with 10 mM phosphate buffer solution at pH7.4. DMEM Salt control composition: ferric nitrate (0.2 µM), calciumchloride (1.8 mM), magnesium sulfate (0.81 mM), potassium chloride (5.3mM), sodium bicarbonate (44 mM), sodium chloride (110 mM) and sodiumphosphate monobasic (0.9 mM). PBS = phosphate buffered saline at pH 7.4,composition: sodium chloride (137 mM), potassium chloride (2.7 mM),disodium phosphate (10 mM), monopotassium phosphate (1.8 mM). ^(a) K_(a)not determined due to intermediate exchange rate on NMR timescaleresulting in complex broad spectra, evidence of binding was indicatedregardless. ^(b) Prepared by dissolution of D-glucono-δ-lactone in 10 mMphosphate buffer, pH 7.4. After 4 h ¹H NMR indicated that the lactonehad hydrolysed to give the acyclic gluconate.

Affinities (K_(d)) were measured in D₂O (NMR) or H₂O (ITC) containingphosphate buffer (10 mM, pH = 7.4) at T = 298 K. N.d. = not determineddue to broadening of NMR signals on addition of substrate

TABLE 5 Summary of binding results for anthracene receptor 90. SubstrateDetermined by NMR K_(a) (M⁻¹) Determined by ITC (kJ mol⁻¹) K_(a) (M⁻¹)ΔG ΔH TΔS D—Glucose 5 ± 3.6% - D—Cellobiose 46 ± 0.9% 38 ± 6.5% -9.0-5.7 3.3 D—cellotriose 950 ± 0.3% 955 ± 1.2% -17.0 -16.5 0.6D—cellotetraose n.d. - D—cellopentaose n.d. - D—maltose 15 ± 11.8% -D—Maltotriose 20 ± 3.3% - Uric Acid - 0

TABLE 6 Measured binding affinities displayed by Receptors 2 to 14towards D-glucose Receptor Binding affinity for D—glucose (K_(a) (M⁻¹))Determined by ITC Determined by¹H- NMR Determined by CD 2 - 13 ± 4 - 35760 ± 269 - - 4 6490 ± 72.6 - - 5 10400 ± 132 - - 6 - - - 7 - 6886 ±190 - 8 4210 ± 73 - - 9 - 2554 ± 96 - 10 - 481 ± 57 - 11 - 2000 1819 ±152 12 0* - - 13 1310 ± 33 - - 14 - 14926 ± 1566 - * No measurablebinding using a 7.1 mM L- or D-glucose solution and a 0.4 mM solution ofReceptor 12.

While specific embodiments of the invention have been described for thepurpose of reference and illustration, various modifications will beapparent to a person skilled in the art without departing from the scopeof the invention as defined by the appended claims.

EMBODIMENTS

1. A compound of Formula (I), or a salt, hydrate or solvate thereof, asshown below:

wherein:

-   bonds b₁ and b₂ are independently selected from a single bond or    double bond;

-   R_(1a), R_(1b), R_(2a) and R_(2b) are independently selected from    hydrogen, carbonyl, (1-8C)alkyl, (3-10C)cycloalkyl, aryl, heteroaryl    and heterocyclyl, each of which, other than hydrogen and carbonyl,    is optionally substituted by one or more substituent groups selected    from (1-4C)alkyl, halo, (1-4C)haloalkyl, (1-4C)haloalkoxy,    (1-4C)alkoxy, (1-4C)alkylamino, amino, cyano, hydroxyl, carboxy,    carbamoyl, sulfamoyl, mercapto and a hydrophilic substituent group;    or

-   R_(1a) and R_(1b) are linked so as to form a group of the formula:

-   

-   and/or R_(2a) and R_(2b) are linked so as to form a group of the    formula:

-   

-   wherein:    -   

    -   denotes the point of attachment;

-   bonds b₁ and b₂ are as described above;

-   Rings A and B are independently selected from aryl, heteroaryl,    heterocyclyl, cycloalkyl and cycloalkenyl;

-   R₁ and R₂ are independently selected from (1-6C)alkyl, halo,    (1-4C)haloalkyl, (1-4C)haloalkoxy, (1-6C)alkoxy, (1-4C)alkylamino,    amino, cyano, hydroxyl, carboxy, carbamoyl, sulfamoyl and mercapto;

-   a and b are integers independently selected from 0 to 2;

-   m and n are integers independently selected from 0 to 2;

-   Z₁ and Z₂ are independently selected from a hydrophilic substituent    group;

-   C and D are independently selected from aryl, heteroaryl,    heterocyclyl, cycloalkyl, cycloalkenyl and a group of the formula:

-   

-   wherein:    -   s, t and v are integers independently selected from 1 or 2;

    -   

    -   denotes the point of attachment;

-   R₃ and R₄ are independently selected from halo, (1-4C)alkyl,    (1-4C)alkoxy, amino, nitro, (1-4C)alkylamino, (1-4C)dialkylamino,    (1-4C)haloalkyl, (1-4C)haloalkoxy, cyano, (2-4C)alkenyl,    (2-4C)alkynyl and a group of the formula:

-   —L¹—Y¹—Q¹ wherein:    -   L¹ is absent or a (1-5C)alkylene optionally substituted by one        or more substituents selected from (1-2C)alkyl and oxo;

    -   Y¹ is absent or selected from a one of the following groups; O,        S, SO, SO₂, N(R_(a)), C(O), C(O)O, OC(O), C(O)N(R_(a)),        N(R_(a))C(O), N(R_(b))C(O)N(R_(a)), N(R_(a))C(O)O,        OC(O)N(R_(a)), S(O)₂N(R_(a)), and N(R_(a))SO₂, wherein R_(a) and        R_(b) are each independently selected from hydrogen and        (1-4C)alkyl; and

    -   Q¹ is hydrogen, (1-8C)alkyl, (2-6C)alkenyl, (2-6C)alkynyl, aryl,        (3-10C)cycloalkyl, (3-10C)cycloalkenyl, heteroaryl and        heterocyclyl; wherein Q¹ is optionally further substituted by        one or more substituent groups independently selected from        (1-4C)alkyl, halo, (1-4C)haloalkyl, (1-4C)haloalkoxy, amino,        (1-4C)aminoalkyl, cyano, hydroxy, carboxy, carbamoyl, sulfamoyl,        mercapto, ureido, oxy, NR_(c)R_(d), OR_(c), C(O)R_(d),        C(O)OR_(c), OC(O)R_(c), C(O)N(R_(d))R_(c), N(R_(d))C(O)R_(c),        S(O)_(y)R_(c) (where y is 0, 1 or 2), SO₂N(R_(d))R_(c),        N(R_(d))SO₂R_(c), Si(R_(e))(R_(d))R_(c) and (CH₂)_(z)NR_(d)R_(c)        (where z is 1, 2 or 3); wherein R_(c), R_(d) and R_(e) are each        independently selected from hydrogen, (1-6C)alkyl and        (3-6C)cycloalkyl; and R_(c) and R_(d) can be linked such that,        together with the nitrogen atom to which they are attached, they        form a 4-7 membered heterocyclic ring which is optionally        substituted by one or more substituents selected from        (1-4C)alkyl, halo, (1-4C)haloalkyl, (1-4C)haloalkoxy,        (1-4C)alkoxy, (1-4C)alkylamino, amino, cyano or hydroxyl; and        wherein two R₃ and/or two R₄ groups taken together may form a        group of the formula:

    -   

    -   wherein:        -   R_(x) is selected from hydrogen and (1-6C)alkyl optionally            substituted by one or more substituent groups selected from            halo, (1-4C)haloalkyl, (1-4C)haloalkoxy, cyano, hydroxy,            sulfamoyl, mercapto, ureido, NR_(f)R_(g), OR_(f), C(O)R_(f),            C(O)OR_(f), OC(O)R_(f), C(O)N(R_(g))R_(f) and            N(R_(g))C(O)R_(f), wherein R_(f) and R_(g) are selected from            hydrogen and (1-4C)alkyl; and        -   the dashed lines represent the points of attachment to C            and/or D;

-   W₁, W₂, W₃ and W₄ are independently selected from CR_(h)R_(i),    wherein R_(h) and R_(i) are selected from hydrogen and (1-2C)alkyl;

-   X₁, X₂, X₃ and X₄ are independently selected from a group of the    formula:

-   

-   wherein:    -   

    -   denotes the point of attachment;

    -   W_(x) is selected from O or NH; and

    -   Q is selected from O, S and NR_(j), wherein R_(j) is selected        from hydrogen, (1-4C)alkyl, aryl, heteroaryl and sulfonyl;

    -   Z₃ and Z₄ are independently selected from a hydrophilic        substituent group;

    -   L is absent or a linker, which optionally bears a hydrophilic        substituent group Z₅;

    -   c and d are integers independently selected from 0 to 4; and

    -   and p are integers independently selected from 0 t2; and        wherein:        -   i) the compound of Formula I is optionally attached to a            displaceable reporter molecule via one or more of the            substituent groups associated with R₁, R₂, R₃, R₄, Z₁, Z₂,            Z₃, Z₄ and/or Z₅; and/or

        -   ii) the compound of Formula I is optionally attached to a            substituent group of Formula A1 shown below at a position            associated with one or more of the substituent groups            R_(1a), R_(1b), R_(2a), R_(2b), R₁, R₂, R₃, R₄, Z₁, Z₂, Z₃,            Z₄ and/or Z₅:

        -   

        -   wherein:            -   X_(2a) is absent or selected from O, S, SO, SO₂,                N(R^(x2)), C(O), C(O)O, OC(O), C(O)N(R^(x2)),                N(R^(x2))C(O), N(R^(x2))C(O)N(R^(x3)), N(R^(x2))C(O)O,                OC(O)N(R^(x2)), S(O)₂N(R^(x2)) and N(R^(x2))SO₂, wherein                R^(x2) and R^(x3) are each independently selected from                hydrogen and (1-4C)alkyl;

            -   L_(2a) is absent or selected from (1-20C)alkylene,                (1-20C)alkylene oxide, (1-20C)alkenyl and                (1-20C)alkynyl, each of which being optionally                substituted by one or more substituents selected from                (1-2C)alkyl, aryl and oxo; and

            -   Z_(2a) is selected from carboxy, carbamoyl, sulphamoyl,                mercapto, amino, azido, (1-4C)alkenyl, (1-4C)alkynyl,                NR^(xc)R^(xd), OR^(xc), ONR^(xc)R^(xd), C(O)X_(a),                C(Q^(z))OR^(xf), N=C=O, NR^(xc)C(O)CH₂X_(b),

            -   C(O)N(R^(xe))NR^(Xc)R^(Xd), S(O)_(y)X_(a) (where y is 0,                1 or 2), SO₂N(R^(xe))NR^(xc)R^(xd),                Si(R^(xg))(R^(xh))R^(xi), S-S-X_(c) an amino acid and

            -   

            -   wherein:

            -   X_(a) is a leaving group (e.g. halo or CF₃);

            -   X_(b) is a halo (e.g. iodo);

            -   X_(c) is an aryl or heteroaryl, optionally substituted                with one or more substituents selected from halo, cyano                and nitro;

            -   R^(xc), R^(xd) and R^(xe) are each independently                selected from hydrogen and (1-6C)alkyl;

            -   R^(xf) is selected from hydrogen and (1-6C)alkyl, or                R^(xf) is a substituent group that renders C(O)OR^(xf),                when taken as a whole, to be an activated ester (e.g a                hydroxysuccinimide ester, a hydroxy-3-sulfo-succinimide                ester or a pentafluorophenyl ester);

            -   Q^(z) is selected from O or ⁺NR^(Q1)R^(Q2), where R^(Q1)                and R^(Q2) are independently selected from hydrogen and                methyl; and

            -   R^(xg), R^(xh) and R^(xi) are each independently                selected from (1-4C)alkyl, hydroxy, halo and                (1-4C)alkoxy;

            -   with the proviso that the compound of Formula I                comprises at least one hydrophilic substituent group                (e.g. Z₁, Z₂, Z₃, Z₄ or Z₅).

2. A compound according to embodiment 1, wherein W₁, W₂, W₃ and W₄ areCH₂.

3. A compound according to embodiments 1 or 2, wherein Q is selectedfrom O or S.

4. A compound according to embodiment 3, wherein Q is O.

5. A compound, salt, hydrate or solvate thereof, according to anypreceding embodiment, wherein the compound has the structural Formula Ibshown below:

wherein, each of R₁, R₂, R₃, R₄, Z₁, Z₂, Z₃, Z₄, a, b, c, d, m, n, o, p,L, C, D and Rings A and B are as defined in any preceding embodiment.

6. A compound according to any preceding embodiment, wherein Rings A andB are independently selected from phenyl, pyridyl, naphthyl andpyrrolidinyl.

7. A compound according to embodiment 6, wherein Rings A and B arephenyl.

8. A compound according to any preceding embodiment, wherein L is alinker selected from a group of the formula:

wherein:

-   

-   denotes the point of attachment;

-   W₅ and W₆ are independently selected from CR_(k)R_(l), wherein R_(k)    and R_(l) are selected from hydrogen and (1-2C)alkyl;

-   X₅ and X₆ are independently selected from a group of the formula:

-   

-   wherein:

-   

-   denotes the point of attachment; and

-   Q₂ is selected from O, S and NR_(m), wherein R_(m), is selected from    hydrogen, (1-4C)alkyl, aryl, heteroaryl and sulfonyl;

-   bond b₃ is a single or double bond;

-   Ring E is selected from aryl, heteroaryl, heterocyclyl, cycloalkyl    and cycloalkenyl;

-   R₅ is selected from (1-4C)alkyl, halo, (1-4C)haloalkyl,    (1-4C)haloalkoxy, (1-4C)alkoxy, (1-4C)alkylamino, amino, cyano,    hydroxyl, carboxy, carbamoyl, sulfamoyl and mercapto;

-   Z₅ is a hydrophilic substituent group;

-   q is an integer from 0 to 2; and

-   e is an integer from 0 to 2.

9. A compound according to embodiment 8, wherein W₅ and W₆ are CH₂.

10. A compound according to any one of embodiments 8 or 9, wherein Q₂ isO.

11. A compound according to any one of embodiments 8 to 10, wherein RingE is selected from phenyl, pyridyl, naphthyl and pyrrolidinyl.

12. A compound according to embodiment 11, wherein Ring E is phenyl.

13. A compound according to any one of embodiments 8 to 12, whereinintegers a and b are 0.

14. A compound according to any one of embodiments 8 to 13, whereinintergers m and n are 1.

15. A compound, salt, hydrate or solvate thereof, according to any oneof embodiments 8 to 14, wherein the compound has the structural formulaId shown below:

wherein, each of C, D, R₃, R₄, Z₁, Z₂, Z₃, Z₄, Z₅, c, d, o and p are asdefined in any preceding embodiment.

16. A compound according any one of embodiments 8 to 15, wherein C and Dare independently selected from phenyl and pyridyl.

17. A compound according to any one of embodiments 8 to 16, wherein Cand D are phenyl.

18. A compound according to any one of embodiments 8 to 17, whereinintegers o and p are 0.

19. A compound, salt, hydrate or solvate thereof, according to any oneof embodiments 8 to 18, wherein the compound has the structural FormulaIe shown below:

wherein:

-   Z¹, Z² and Z⁵ are as defined in embodiment 1; and-   R^(3a), R^(3b), R^(3c), R^(4a), R^(4b) and R^(4c) are independently    selected from hydrogen, halo, (1-4C)alkyl, (1-4C)alkoxy, amino,    nitro, (1-4C)alkylamino, (1-4C)dialkylamino, (1-4C)haloalkyl,    (1-4C)haloalkoxy, cyano, (2-4C)alkenyl, (2-4C)alkynyl and a group of    the formula:-   —L^(1a)—Y^(1a)—Q^(1a) wherein:    -   L^(1a) is absent or (1-2C)alkylene optionally substituted by one        or more substituents selected from (1-2C)alkyl or oxo;    -   Y^(1a) is absent or selected from a one of the following groups;        O, S, SO, SO₂, N(R_(n)), C(O), C(O)O, OC(O), C(O)N(R_(n)) and        N(R_(n))C(O), wherein R_(n) is selected from hydrogen and        (1-4C)alkyl; and    -   Q^(1a) is hydrogen, (1-8C)alkyl, (2-6C)alkenyl, (2-6C)alkynyl,        aryl, (3-10C)cycloalkyl, (3-10C)cycloalkenyl, heteroaryl and        heterocyclyl; wherein Q^(1a) is optionally further substituted        by one or more substituent groups independently selected from        (1-4C)alkyl, halo, (1-4C)haloalkyl, (1-4C)haloalkoxy, amino,        (1-4C)aminoalkyl, cyano, hydroxy, carboxy, carbamoyl, sulfamoyl,        mercapto, ureido, oxy, NR_(o)R_(p), OR_(o), C(O)R_(o),        C(O)OR_(o), OC(O)R_(o), C(O)N(R_(p))R_(o), N(R_(p))C(O)R_(o),        S(O)_(y1)R_(o) (where y₁ is 0, 1 or 2), SO₂N(R_(p))R_(o),        N(R_(p))SO₂R_(o), Si(R_(q))(R_(p))R_(o) and        (CH₂)_(z1)NR_(o)R_(p) (where z₁ is 1, 2 or 3); wherein R_(o),        R_(p) and R_(q) are each independently selected from hydrogen        and (1-6C)alkyl.

20. A compound according to embodiments 19, wherein R^(3a), R^(3b),R^(3c), R^(4a), R^(4b) and R^(4c) are independently selected fromhydrogen, halo, (1-4C)alkyl, (1-4C)alkoxy, amino, nitro,(1-4C)alkylamino, (1-4C)dialkylamino, (1-4C)haloalkyl, (1-4C)haloalkoxy,cyano, (2-4C)alkenyl, (2-4C)alkynyl and a group of the formula:

wherein:

-   L^(1a) is absent or (1-2C)alkylene;-   Y^(1a) is absent or selected from a one of the following groups; O,    S, SO, SO₂, N(R_(l)), C(O), C(O)O, OC(O), C(O)N(R_(n)) and    N(R_(n))C(O), wherein R_(n) is selected from hydrogen and    (1-4C)alkyl; and-   Q^(1a) is hydrogen, (1-8C)alkyl, (2-6C)alkenyl, (2-6C)alkynyl, aryl,    (3-10C)cycloalkyl, (3-10C)cycloalkenyl, heteroaryl and heterocyclyl;    wherein Q^(1a) is optionally further substituted by one or more    substituent groups independently selected from (1-4C)alkyl, halo,    (1-4C)haloalkyl, (1-4C)haloalkoxy, amino, (1-4C)aminoalkyl, cyano,    hydroxy, carboxy, carbamoyl, sulfamoyl and mercapto.

21. A compound according any one of embodiments 19 or 20, whereinR^(3a), R^(3b), R^(3c), R^(4a), R^(4b) and R^(4c) are independentlyselected from hydrogen, halo, (1-4C)alkyl, (1-4C)alkoxy, amino, nitro,(1-4C)alkylamino, (1-4C)dialkylamino, (1-4C)haloalkyl, (1-4C)haloalkoxy,cyano, (2-4C)alkenyl and (2-4C)alkynyl.

22. A compound according to embodiments 1 to 7, wherein L is absent.

23. A compound according embodiment 22, wherein C and D areindependently selected from naphthenyl or anthracenyl.

24. A compound according embodiment 23, wherein C and D are anthracenyl.

25. A compound according to any one of embodiments 22 to 24, wherein thecompound has the structural Formula Ig, shown below:

wherein each of R₁, R₂, R₃, R₄, Z₁, Z₂, Z₃, Z₄, a, b, c, d, m, n, o andp are as defined in embodiment 1.

26. A compound according to any one of embodiments 22 to 25, whereinintegers m and n are 0.

27. A compound according to any one of embodiments 22 to 26, whereinintergers a and b are 0.

28. A compound according to any preceding embodiment, wherein Z₁, Z₂, Z₃and Z₄ and Z₅ are independently selected from a hydrophilic substituentgroup comprising one or more hydrophilic functional groups selected fromcarboxylic acids, carboxylate ions, carboxylate esters, hydroxyl,amines, amides, ethers, ketone and aldehyde groups, nitro groups,sulphates, sulphonates, phosphates, phosphonates, and combinationsthereof.

29. A compound according to any one of embodiments 1 to 27 embodiment,wherein Z₁, Z₂, Z₃ and Z₄ and Z₅ are independently selected from ahydrophilic substituent group, wherein said hydrophilic substituentgroup is a hydrophilic polymer or hydrophilic dendritic group.

30. A compound according to any one of embodiments 1 to 27 embodiment,wherein Z₁, Z₂, Z₃ and Z₄ and Z₅ are independently selected from ahydrophilic polymer or a dendritic group comprising between 1 and 5generations of building units and a terminal functional group T₁, andwherein each building unit is independently selected from a group ofFormula A:

wherein:

-   L² is selected from O, C(O), C(O)O, OC(O), C(O)N(R_(r)),    N(R_(r))C(O), N(R_(s))C(O)N(R_(r)), N(R_(r))C(O)O, OC(O)N(R_(r)),    S(O)₂N(R_(r)), and N(R_(r))SO₂, wherein R_(r) and R_(s) are each    independently selected from hydrogen and (1-4C)alkyl;

-   L^(2a) is a bond or a (1-4C)alkylene;

-   V is absent or a group of the formula:

-   

-   

-   wherein:    -   V₁, V₂, V₃, V₄ and V₅ are independently selected from a        (1-6C)alkylene optionally interrupted by one or more groups        selected from O, S and NR_(t), wherein R_(t) is selected from        hydrogen and (1-2C)alkyl;

    -   #denotes the point of attachment to one of Rings A, B, C, D or        E;

    -   

    -   denotes the point of attachment to either another group of        Formula A or a terminal functional group T₁; and

    -   the terminal functional group T₁ is selected from OH,        C(O)OM_(x), C(O)OR_(u) and C(O)NHR_(u), wherein R_(u) is        selected from hydrogen, (1-4C)alkyl, (1-4C)alkoxy,        hydroxy(1-4C)alkyl, (2-4C)alkenyl, (2-4C)alkynyl, ethylene        gylycol and polyethylene glycol, and wherein M_(x) is a cation        (e.g. Na, Li, NH₄).

31. A compound according to embodiment 30, wherein the dendritic groupcomprises between 1 and 4 generations of building units and a terminalfunctional group T₁, and wherein each building unit is independentlyselected from a group of Formula A:

wherein:

-   L² is selected from O, C(O), C(O)O and C(O)N(R_(r)), wherein R_(r)    is selected from hydrogen and (1-4C)alkyl;

-   L^(2a) is a bond or a (1-4C)alkylene;

-   V is absent or a group of the formula:

-   

-   wherein:    -   V₁, V₂, and V₃ are independently selected from a (1-6C)alkylene        optionally interrupted by one or more groups selected from        oxygen atoms;

    -   #denotes the point of attachment to one of Rings A, B, C, D or        E;

    -   

    -   denotes the point of attachment to either another group of        Formula A or a terminal functional group T₁; and

    -   the terminal functional group T₁ is selected from OH,        C(O)OM_(x), C(O)OR_(u) and C(O)NHR_(u), wherein R_(u) is        selected from hydrogen, (1-4C)alkoxy and hydroxy(1-4C)alkyl,        wherein M_(x) is a cation (e.g. Na, Li, NH₄).

32. A compound according to any one of embodiments 30 to 31, wherein thedendritic group comprises between 1 and 3 generations of building unitsand a terminal functional group T₁, and wherein each building unit isindependently selected from a group of Formula A:

wherein:

-   L² is C(O)N(R_(r)), wherein R_(r) is selected from hydrogen and    (1-4C)alkyl;

-   L^(2a) is a bond or a (1-2C)alkylene;

-   V is a group of the formula:

-   

-   wherein:    -   V₁, V₂, and V₃ are independently selected from a (1-4C)alkylene        optionally interrupted by one or more groups selected from        oxygen atoms;

    -   #denotes the point of attachment to one of Rings A, B, C, D or        E;

    -   

    -   denotes the point of attachment to either another group of        Formula A or a terminal functional group T₁; and

    -   the terminal functional group T₁ is selected is C(O)OM_(x),        wherein M_(x) is a cation (e.g. Na, Li, NH₄).

33. A compound selected from any one of the following:

34. A compound of the formula shown below:

-   wherein each of Z₁, Z₂ and Z₅ is a group of the formula:

-   

-   wherein

-   

-   denotes the point of attachment.

35. A compound of the formula shown below:

-   wherein each of Z₁ and Z₂ is a group of the formula:

-   

-   wherein

-   

-   denotes the point of attachment.

36. A compound of the formula shown below:

-   wherein each of Z₃, and Z₄ is a group of the formula:

-   

-   wherein

-   

-   denotes the point of attachment.

37. A compound according to any one of the preceding embodiments, whichis immobilised on or in a solid or semi-solid support.

38. A compound according to embodiment 37, wherein the solid orsemi-solid support is a polymeric matrix and/or a gel, such as ahydrogel.

39. A compound according to embodiment 38, wherein the polymeric matrixand/or gel is a polymer selected from cross-linked polyethylene glycoland/or polyacrylamide.

40. A compound according to embodiments 38 or 39, wherein the compoundsare chemically linked to the polymeric matrix and/or gel.

41. A compound according to embodiments 38 or 39, wherein the compoundsare physically incorporated within the polymeric matrix and/or gel vianon-covalent interactions.

42. A complex comprising a compound according to any one of embodiments1 to 41 in association with a displaceable reporter molecule.

43. A composition comprising a compound according to any one ofembodiments 1 to 41 and a displaceable reporter molecule.

44. A complex according to embodiment 42 or a composition according toembodiment 43, wherein the displaceable reporter molecule is an aromaticmolecule or dye.

45. A complex or composition according to embodiments 44, wherein thedisplaceable reporter molecule is a fluorescent aromatic molecule.

46. A complex or composition according to any one of embodiments 44 or45, wherein the compound is as defined in embodiment 15.

47. A saccharide detection device comprising a complex according to anyone of embodiments 41 or 43 to 45, a composition according to any one ofembodiments 43 to 46, or a compound according to any one of embodiments1 to 41.

48. Use of a complex according to any one of embodiments 42 or 44 to 46,a composition according to any one of embodiments 43 to 46, a compoundaccording to to any one of embodiments 1 to 40, or a saccharidedetection device according to embodiment 47, for detecting a targetsaccharide in an aqueous environment.

49. Use according to embodiment 48, wherein the target saccharide isglucose.

50. Use according to any one of embodiments 48 or 49, wherein theaqueous environment is blood or blood plasma.

51. Use according to any one of embodiments 48 to 49, wherein theaqueous environment is a fermentation medium.

52. Use of a complex according to any one of embodiments 42 or 44 to 46,a composition according to any one of embodiments 43 to 46, a compoundaccording to any one of embodiments 1 to 40, or a saccharide detectiondevice according to embodiment 47, for the diagnosis of a conditionwhich results in, or is otherwise associated with, an abnormalconcentration of, and/or a change in the concentration of, a targetsaccharide.

53. Use according to embodiment 52, wherein the condition is diabetes.

54. Use of a compound according to any one of embodiments 1 to 41, in aglucose responsive insulin based system.

55. A complex comprising a compound according to any one of embodiments1 to 41, covalently attached to insulin.

1. A compound of Formula (I), or a salt, hydrate or solvate thereof, asshown below:

wherein: bonds b₁ and b₂ are independently selected from a single bondor double bond; R_(1a), R_(1b), R_(2a) and R_(2b) are independentlyselected from hydrogen, carbonyl, (1-8C)alkyl, (3-10C)cycloalkyl, aryl,heteroaryl and heterocyclyl, each of which, other than hydrogen andcarbonyl, is optionally substituted by one or more substituent groupsselected from (1-4C)alkyl, halo, (1-4C)haloalkyl, (1-4C)haloalkoxy,(1-4C)alkoxy, (1-4C)alkylamino, amino, cyano, hydroxyl, carboxy,carbamoyl, sulfamoyl, mercapto and a hydrophilic substituent group; orR_(1a) and R_(1b) are linked so as to form a group of the formula:

and/or R_(2a) and R_(2b) are linked so as to form a group of theformula:

wherein:

denotes the point of attachment; bonds b₁ and b₂ are as described above;Rings A and B are independently selected from aryl, heteroaryl,heterocyclyl, cycloalkyl and cycloalkenyl; R₁ and R₂ are independentlyselected from (1-6C)alkyl, halo, (1-4C)haloalkyl, (1-4C)haloalkoxy,(1-6C)alkoxy, (1-4C)alkylamino, amino, cyano, hydroxyl, carboxy,carbamoyl, sulfamoyl and mercapto; a and b are integers independentlyselected from 0 to 2; m and n are integers independently selected from 0to 2; Z₁ and Z₂ are independently selected from a hydrophilicsubstituent group; C and D are independently selected from aryl,heteroaryl, heterocyclyl, cycloalkyl, cycloalkenyl and a group of theformula:

wherein: s, t and v are integers independently selected from 1 or 2;

denotes the point of attachment; R₃ and R₄ are independently selectedfrom halo, (1-4C)alkyl, (1-4C)alkoxy, amino, nitro, (1-4C)alkylamino,(1-4C)dialkylamino, (1-4C)haloalkyl, (1-4C)haloalkoxy, cyano,(2-4C)alkenyl, (2-4C)alkynyl and a group of the formula: —L¹—Y¹—Q¹wherein: L¹ is absent or a (1-5C)alkylene optionally substituted by oneor more substituents selected from (1-2C)alkyl and oxo; Y¹ is absent orselected from a one of the following groups; O, S, SO, SO₂, N(R_(a)),C(O), C(O)O, OC(O), C(O)N(R_(a)), N(R_(a))C(O), N(R_(b))C(O)N(R_(a)),N(R_(a))C(O)O, OC(O)N(R_(a)), S(O)₂N(R_(a)), and N(R_(a))SO₂, whereinR_(a) and R_(b) are each independently selected from hydrogen and(1-4C)alkyl; and Q¹ is hydrogen, (1-8C)alkyl, (2-6C)alkenyl,(2-6C)alkynyl, aryl, (3-10C)cycloalkyl, (3-10C)cycloalkenyl, heteroaryland heterocyclyl; wherein Q¹ is optionally further substituted by one ormore substituent groups independently selected from (1-4C)alkyl, halo,(1-4C)haloalkyl, (1-4C)haloalkoxy, amino, (1-4C)aminoalkyl, cyano,hydroxy, carboxy, carbamoyl, sulfamoyl, mercapto, ureido, oxy,NR_(c)R_(d), OR_(c,), C(O)R_(d), C(O)OR_(c), OC(O)R_(c),C(O)N(R_(d))R_(c), N(R’_(d))C(O)R_(c), S(O)_(y)R_(c) (where y is 0, 1 or2), SO₂N(R_(d))R_(c), N(R_(d))SO₂R_(c), Si(R_(e))(R_(d))R_(c) and(CH₂)_(z)NR_(d)R_(c) (where z is 1, 2 or 3); wherein R_(c), R_(d) andR_(e) are each independently selected from hydrogen, (1-6C)alkyl and(3-6C)cycloalkyl; and R_(c) and R_(d) can be linked such that, togetherwith the nitrogen atom to which they are attached, they form a 4-7membered heterocyclic ring which is optionally substituted by one ormore substituents selected from (1-4C)alkyl, halo, (1-4C)haloalkyl,(1-4C)haloalkoxy, (1-4C)alkoxy, (1-4C)alkylamino, amino, cyano orhydroxyl; and wherein two R₃ and/or two R₄ groups taken together mayform a group of the formula:

wherein: R_(x) is selected from hydrogen and (1-6C)alkyl optionallysubstituted by one or more substituent groups selected from halo,(1-4C)haloalkyl, (1-4C)haloalkoxy, cyano, hydroxy, sulfamoyl, mercapto,ureido, NR_(f)R_(g), OR_(f), C(O)R_(f), C(O)OR_(f), OC(O)R_(f),C(O)N(R_(g))R_(f) and N(R_(g))C(O)R_(f), wherein R_(f) and R_(g) areselected from hydrogen and (1-4C)alkyl; and the dashed lines representthe points of attachment to C and/or D; W₁, W₂, W₃ and W₄ areindependently selected from CR_(h)R_(i), wherein R_(h) and R_(i) areselected from hydrogen and (1-2C)alkyl; X₁, X₂, X₃ and X₄ areindependently selected from a group of the formula:

wherein:

denotes the point of attachment; W_(x) is selected from O or NH; and Qis selected from O, S and NR_(j), wherein R_(j) is selected fromhydrogen, (1-4C)alkyl, aryl, heteroaryl and sulfonyl; Z₃ and Z₄ areindependently selected from a hydrophilic substituent group; L is absentor a linker, which optionally bears a hydrophilic substituent group Z₅;c and d are integers independently selected from 0 to 4; and and p areintegers independently selected from 0 t2; and wherein: iii) thecompound of Formula I is optionally attached to a displaceable reportermolecule via one or more of the substituent groups associated with R₁,R₂, R₃, R₄, Z₁, Z₂, Z₃, Z₄ and/or Z₅; and/or iv) the compound of FormulaI is optionally attached to a substituent group of Formula A1 shownbelow at a position associated with one or more of the substituentgroups R_(1a), R_(1b), R_(2a), R_(2b), R₁, R₂, R₃, R₄, Z₁, Z₂, Z₃, Z₄and/or Z₅:

wherein: X_(2a) is absent or selected from O, S, SO, SO₂, N(R^(x2)),C(O), C(O)O, OC(O), C(O)N(R^(x2)), N(R^(x2))C(O),N(R^(x2))C(O)N(R^(x3)), N(R^(x2))C(O)O, OC(O)N(R^(x2)), S(O)₂N(R^(x2))and N(R^(x2))SO₂, wherein R^(x2) and R^(x3) are each independentlyselected from hydrogen and (1-4C)alkyl; L_(2a) is absent or selectedfrom (1-20C)alkylene, (1-20C)alkylene oxide, (1-20C)alkenyl and(1-20C)alkynyl, each of which being optionally substituted by one ormore substituents selected from (1-2C)alkyl, aryl and oxo; and Z_(2a) isselected from carboxy, carbamoyl, sulphamoyl, mercapto, amino, azido,(1-4C)alkenyl, (1-4C)alkynyl, NR^(xc)R^(xd), OR^(xc), ONR^(xc)R^(xd),C(O)X_(a), C(Q^(z))OR^(xf), N=C=O, NR^(xc)C(O)CH₂X_(b),C(O)N(R^(xe))NR^(Xc)R^(Xd), S(O)_(y)X_(a) (where y is 0, 1 or 2),SO₂N(R^(xe))NR^(xc)R^(xd), Si(R^(xg))(R^(xh))R^(xi), S—S—X_(c) an aminoacid and

wherein: X_(a) is a leaving group (e.g. halo or CF₃); X_(b) is a halo(e.g. iodo); X_(c) is an aryl or heteroaryl, optionally substituted withone or more substituents selected from halo, cyano and nitro; R^(xc),R^(xd) and R^(xe) are each independently selected from hydrogen and(1-6C)alkyl; R^(xf) is selected from hydrogen and (1-6C)alkyl, or R^(xf)is a substituent group that renders C(O)OR^(xf), when taken as a whole,to be an activated ester (e.g a hydroxysuccinimide ester, ahydroxy-3-sulfo-succinimide ester or a pentafluorophenyl ester); Q^(z)is selected from O or ⁺NR^(Q1) R^(Q2), where R^(Q1) and R^(Q2) areindependently selected from hydrogen and methyl; and R^(xg), R^(xh) andR^(xi) are each independently selected from (1-4C)alkyl, hydroxy, haloand (1-4C)alkoxy; with the proviso that the compound of Formula Icomprises at least one hydrophilic substituent group (e.g. Z₁, Z₂, Z₃,Z₄ or Z₅).